Researcher Profile and Settings

Affiliation

• Faculty of Dental Medicine　Division of Dental Medicine　Department of Pathobiological Science

Job Title

• Professor

Degree

• 1998 PhD (Hokkaido Univ. Dental School)

URL

https://www.den.hokudai.ac.jp/vascular-biol-pathol/
http://www.den.hokudai.ac.jp/vascular-biology/

J-Global ID

• 200901013920492498

Research Interests

• 腫瘍学   血管新生   細胞生物学   angiogenesis   Cell Biology   Oncology   

Research Areas

• Life sciences / Surgical dentistry
• Life sciences / Experimental pathology
• Life sciences / Pathobiochemistry

Educational Organization

•  Bachelor's degree program　School of Dental Medicine
•  Doctoral (PhD) degree program　Graduate school of Dental Medicine

Academic & Professional Experience

• 2018/05 - Today Hokkaido University Vascular Biology and Molecular Pathology,Graduate School of Dental Medicine rofessor
• 2014/04 - 2018/04 Hokkaido University Institute for Genetic Medicine
• 2009/04 - 2014/03 北海道大学大学院歯学研究科口腔病態学講座血管生物学教室・特任准教授
• 2007/04 - 2009/03 北海道大学大学院歯学研究科口腔病態学講座口腔病理病態学・助教
• 2005/04 - 2007/03 Hokkaido University Graduate School of Dental Medicine
• 2005 - 2007 Research Associate
• 2004/01 - 2005/03 ハーバード大医学部小児病院 Vascular Research Program・助手
• 2004 - 2005 Research Associate
• 2001/07 - 2003/12 ハーバード大医学部小児病院Vascular Research Program・助手
• 2001 - 2003 Researcher
• 2000/09 - 2001/01 Hokkaido University School of Dental Medicine
• 2000 - 2001 Medical Staff
• 1998/04 - 2000/03 日本学術振興会特別研究員 日本学術振興会特別研究員（PD）
• 1998 - 2000 Postdoctoral Fellowships of Japan Society for the Promotion of Science
• 1994/04 - 1995/03 札幌鉄道病院口腔外科・医師
• 1994 - 1995 Medical Staff
• 1992/07 - 1994/03 Hokkaido University School of Dental Medicine
• 1992 - 1994 Medical Staff

Education

• 1995/04 - 1998/03  Hokkaido University  Graduate School of Dental Medicine
•        - 1998  Hokkaido University  Graduate School, Division of Dental Research  PhD
• 1986/04 - 1992/03  Hokkaido University  School of Dental Medicine
•        - 1992  Hokkaido University  Faculty of Dentistry

Association Memberships

• Council Member Science council Japan   Science council Japan   日本がん免疫学会   日本臨床口腔病理学会   日本細胞外小胞学会   THE MOLECULAR BIOLOGY SOCIETY OF JAPAN   THE SOCIETY OF JAPANESE WOMEN SCIENTISTS   日本がん転移学会   JAPAN SOCIETY FOR CELL BIOLOGY   歯科基礎医学会   JAPANESE SOCIETY OF ORAL AND MAXILLOFACIAL SURGEONS   JAPANESE STOMATOLOGICAL SOCIETY   THE JAPANESE CANCER ASSOCIATION   アメリカ癌学会   日本血管生物医学会   THE JAPANESE SOCIETY OF PATHOLOGY   Japanese Association of PathologyJapanese association of Pathology   American Association of cancer research   北海道歯学会   

Research Activities

Published Papers

• Viral uptake and pathophysiology of the lung endothelial cells in age‐associated severe SARS‐CoV‐2 infection models

  Takuya Tsumita, Ryo Takeda, Nako Maishi, Yasuhiro Hida, Michihito Sasaki, Yasuko Orba, Akihiko Sato, Shinsuke Toba, Wataru Ito, Takahito Teshirogi, Yuya Sakurai, Tomohiro Iba, Hisamichi Naito, Hitoshi Ando, Haruhisa Watanabe, Amane Mizuno, Toshiki Nakanishi, Aya Matsuda, Ren Zixiao, Ji‐Won Lee, Tadahiro Iimura, Hirofumi Sawa, Kyoko Hida

  Aging Cell e14050 1474-9718 2023/12/14 [Refereed][Not invited]
   

  Abstract Thrombosis is the major cause of death in severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection, and the pathology of vascular endothelial cells (ECs) has received much attention. Although there is evidence of the infection of ECs in human autopsy tissues, their detailed pathophysiology remains unclear due to the lack of animal model to study it. We used a mouse‐adapted SARS‐CoV‐2 virus strain in young and mid‐aged mice. Only mid‐aged mice developed fatal pneumonia with thrombosis. Pulmonary ECs were isolated from these infected mice and RNA‐Seq was performed. The pulmonary EC transcriptome revealed that significantly higher levels of viral genes were detected in ECs from mid‐aged mice with upregulation of viral response genes such as DDX58 and IRF7. In addition, the thrombogenesis‐related genes encoding PLAT, PF4, F3 PAI‐1, and P‐selectin were upregulated. In addition, the inflammation‐related molecules such as CXCL2 and CXCL10 were upregulated in the mid‐aged ECs upon viral infection. Our mouse model demonstrated that SARS‐CoV‐2 virus entry into aged vascular ECs upregulated thrombogenesis and inflammation‐related genes and led to fatal pneumonia with thrombosis. Current results of EC transcriptome showed that EC uptake virus and become thrombogenic by activating neutrophils and platelets in the aged mice, suggesting age‐associated EC response as a novel finding in human severe COVID‐19.
• Oral bacterium Streptococcus mutans promotes tumor metastasis through thrombosis formation

  Li Yu, Yuying Hong, Nako Maishi, Aya Yanagawa Matsuda, Yasuhiro Hida, Akira Hasebe, Yoshimasa Kitagawa, Kyoko Hida

  Cancer Science 115 (2) 648 - 659 1347-9032 2023/12/14 [Refereed]
   

  Abstract Thrombosis is a well‐known cardiovascular disease (CVD) complication that has caused death in many patients with cancer. Oral bacteria have been reported to contribute to systemic diseases, including CVDs, and tumor metastasis. However, whether oral bacteria‐induced thrombosis induces tumor metastasis remains poorly understood. In this study, the cariogenic oral bacterium Streptococcus mutans was used to examine thrombosis in vitro and in vivo. Investigation of tumor metastasis to the lungs was undertaken by intravenous S. mutans implantation using a murine breast cancer metastasis model. The results indicated that platelet activation, aggregation, and coagulation were significantly altered in S. mutans‐stimulated endothelial cells (ECs), with elevated neutrophil migration, thereby inducing thrombosis formation. Streptococcus mutans stimulation significantly enhances platelet and tumor cell adhesion to the inflamed ECs. Furthermore, S. mutans‐induced pulmonary thrombosis promotes breast cancer cell metastasis to the lungs in vivo, which can be reduced by using aspirin, an antiplatelet drug. Our findings indicate that oral bacteria promote tumor metastasis through thrombosis formation. Oral health management is important to prevent CVDs, tumor metastasis, and their associated death.
• Low-dose metronomic cisplatin as an antiangiogenic and anti-inflammatory strategy for cancer

  Hiroshi Kikuchi, Nako Maishi, Li Yu, Zi Jia, Cong Li, Masumi Sato, Ryo Takeda, Keita Ishizuka, Yasuhiro Hida, Nobuo Shinohara, Kyoko Hida

  British Journal of Cancer 0007-0920 2023/12/01 [Refereed]
  
• Significant association of Yamamoto-Kohama classification and pathological depth of invasion with cervical lymph node metastasis in early-stage tongue squamous cell carcinoma (Stage I/II)

  Wataru Kakuguchi, Yuichi Ashikaga, Aya Yanagawa-Matsuda, Kazuyo Kuribayashi, Saki Shinohara, Naohiro Ogawa, Nako Maishi, Kyoko Hida, Chih-Yuan Fang, Yoichi Ohiro

  Journal of Dental Sciences 18 (4) 1663 - 1668 1991-7902 2023/02
• Bovine Lactoferrin Suppresses Tumor Angiogenesis through NF-κB Pathway Inhibition by Binding to TRAF6

  Nurina Febriyanti Ayuningtyas, Chanbora Chea, Toshinori Ando, Karina Erda Saninggar, Keiji Tanimoto, Toshihiro Inubushi, Nako Maishi, Kyoko Hida, Masanobu Shindoh, Mutsumi Miyauchi, Takashi Takata

  Pharmaceutics 15 (1) 165 - 165 2023/01/03 

  Tumor angiogenesis is essential for tumor progression. The inhibition of tumor angiogenesis is a promising therapy for tumors. Bovine lactoferrin (bLF) has been reported as an anti-tumor agent. However, bLF effects on tumor angiogenesis are not well demonstrated. This study evaluated the inhibitory effects of bLF on tumor angiogenesis in vivo and in vitro. Herein, tumor endothelial cells (TECs) and normal endothelial cells (NECs) were used. Proliferation, migration, tube formation assays, RT-PCR, flow cytometry, Western blotting, siRNA experiments and immunoprecipitation were conducted to clarify the mechanisms of bLF-induced effects. CD-31 immunoexpression was examined in tumor tissues of oral squamous cell carcinoma mouse models with or without Liposomal bLF (LbLF)-administration. We confirmed that bLF inhibited proliferation/migration/tube formation and increased apoptosis in TECs but not NECs. TNF receptor-associated factor 6 (TRAF6), p-p65, hypoxia inducible factor-α (HIF-1α) and vascular endothelial growth factor (VEGF) were highly expressed in TECs. In TECs, bLF markedly downregulated VEGF-A, VEGF receptor (VEGFR) and HIF-1α via the inhibition of p-p65 through binding with TRAF6. Since NECs slightly expressed p-p65, bLF–TRAF-6 binding could not induce detectable changes. Moreover, orally administrated LbLF decreased CD31-positive microvascular density only in TECs. Hence, bLF specifically suppressed tumor angiogenesis through p-p65 inhibition by binding to TRAF6 and suppressing HIF-1α activation followed by VEGF/VEGFR down-regulation. Collectively, bLF can be an anti-angiogenic agent for tumors.
• miR-1246 in tumor extracellular vesicles promotes metastasis via increased tumor cell adhesion and endothelial cell barrier destruction

  Morimoto M, Maishi N, Tsumita T, Alam MT, Kikuchi H, Hida Y, Yoshioka Y, Ochiya T, Annan DA, Takeda R, Kitagawa Y, Hida K

  Front. Oncol 13 973871 - 973871 2023 [Refereed]
   

  BACKGROUND: Tumor blood vessels play a key role in tumor metastasis. We have previously reported that tumor endothelial cells (TECs) exhibit abnormalities compared to normal endothelial cells. However, it is unclear how TECs acquire these abnormalities. Tumor cells secrete extracellular vesicles (EVs) to create a suitable environment for themselves. We have previously identified miR-1246 to be more abundant in high metastatic melanoma EVs than in low metastatic melanoma EVs. In the current study, we focused on miR-1246 as primarily responsible for acquiring abnormalities in TECs and examined whether the alteration of endothelial cell (EC) character by miR-1246 promotes cancer metastasis. METHODS: We analyzed the effect of miR-1246 in metastatic melanoma, A375SM-EVs, in vivo metastasis. The role of tumor EV-miR-1246 in the adhesion between ECs and tumor cells and the EC barrier was addressed. Changes in the expression of adhesion molecule and endothelial permeability were examined. RESULTS: Intravenous administration of A375SM-EVs induced tumor cell colonization in the lung resulting in lung metastasis. In contrast, miR-1246 knockdown in A375SM decreased lung metastasis in vivo. miR-1246 transfection in ECs increased the expression of adhesion molecule ICAM-1 via activation of STAT3, followed by increased tumor cell adhesion to ECs. Furthermore, the expression of VE-Cadherin was downregulated in miR-1246 overexpressed EC. A375SM-EV treatment enhanced endothelial permeability. VE-Cadherin was validated as the potential target gene of miR-1246 via the target gene prediction database and 3' UTR assay. CONCLUSION: miR-1246 in high metastatic tumor EVs promotes lung metastasis by inducing the adhesion of tumor cells to ECs and destroying the EC barrier.
• Angiogenic inhibitor pre-administration improves the therapeutic effects of immunotherapy

  Sato M, Maishi N, Hida Y, Yanagawa-Matsuda A, Alam MT, Sakakibara-Konishi J, Nam JM, Onodera Y, Konno S, Hida K

  Cancer Med 2023 [Refereed]
   

  In lung cancer, immune checkpoint inhibitors (ICIs) are often inadequate for tumor growth inhibition. Angiogenic inhibitors (AIs) are required to normalize tumor vasculature for improved immune cell infiltration. However, in clinical practice, ICIs and cytotoxic antineoplastic agents are simultaneously administered with an AI when tumor vessels are abnormal. Therefore, we examined the effects of pre-administering an AI for lung cancer immunotherapy in a mouse lung cancer model. Using DC101, an anti-vascular endothelial growth factor receptor 2 (VEGFR2) monoclonal antibody, a murine subcutaneous Lewis lung cancer (LLC) model was used to determine the timing of vascular normalization. Microvessel density (MVD), pericyte coverage, tissue hypoxia, and CD8-positive cell infiltration were analyzed. The effects of an ICI and paclitaxel after DC101 pre-administration were investigated. On Day 3, increased pericyte coverage and alleviated tumor hypoxia represented the highest vascular normalization. CD8+ T-cell infiltration was also highest on Day 3. When combined with an ICI, DC101 pre-administration significantly reduced PD-L1 expression. When combined with an ICI and paclitaxel, only DC101 pre-administration significantly inhibited tumor growth, but simultaneous administration did not. AI pre-administration, and not simultaneous administration, may increase the therapeutic effects of ICIs due to improved immune cell infiltration.
• Stroma biglycan expression can be a prognostic factor in prostate cancers

  Jun Furumido, Nako Maishi, Aya Yanagawa‐Matsuda, Hiroshi Kikuchi, Ryuji Matsumoto, Takahiro Osawa, Takashige Abe, Yoshihiro Matsuno, Nobuo Shinohara, Yasuhiro Hida, Kyoko Hida

  International Journal of Urology 30 (2) 147 - 154 0919-8172 2022/10/28 [Refereed]
   

  OBJECTIVES: This study analyzes the relationship between biglycan expression in prostate cancer and clinicopathological parameters to clarify the potential link between biglycan and prognosis and progression to castration-resistant prostate cancer (CRPC). METHODS: We retrospectively analyzed 60 cases of prostate cancer patients who underwent robot-assisted laparoscopic radical prostatectomy in Hokkaido University Hospital. RESULTS: Biglycan was expressed in the tumor stroma but not in tumor cells. There was no significant relationship with biochemical recurrence (p = 0.5237), but the expression of biglycan was 36.1% in the group with progression to CRPC. This indicates a significant relationship with progression to CRPC (p = 0.0182). Furthermore, the expression of biglycan-positive blood vessels was significantly higher (15.9%) in the group with biochemical recurrence than in the group without biochemical recurrence (8.5%) (p = 0.0169). The biglycan-positive vessels were 28.6% in the group with progression to CRPC, which was significantly higher than that in the group without progression to CRPC (p < 0.0001). CONCLUSION: This is the first study to show that stroma biglycan is a useful prognostic factor for prostate cancer.
• The oxidized‐LDL/LOX‐1 axis in tumor endothelial cells enhances metastasis by recruiting neutrophils and cancer cells

  Takuya Tsumita, Nako Maishi, Dorcas Akuba‐Muhyia Annan, Mohammad Alam Towfik, Aya Matsuda, Yasuhito Onodera, Jin‐Min Nam, Yasuhiro Hida, Kyoko Hida

  International Journal of Cancer 151 (6) 944 - 956 0020-7136 2022/09/15 [Refereed][Not invited]
   

  Epidemiological relationships between cancer and cardiovascular diseases have been reported, but a molecular basis remains unclear. Some proteoglycans that strongly bind low-density-lipoprotein (LDL) are abundant both in atherosclerotic regions and in high metastatic-tumor tissue. LDL retention is crucial for the initiation of atherosclerosis, although its contribution to malignancy of cancer is not known. In our study, we show the importance of the accumulation of LDL in tumor metastasis. We demonstrated that high metastatic-tumor tissue contains high amounts of LDL and forms more oxidized LDL (ox-LDL). Interestingly, lectin-like ox-LDL receptor 1 (LOX-1), a receptor for ox-LDL and a recognized key molecule for cardiovascular diseases, was highly expressed in tumor endothelial cells (TECs). Neutrophils are important for ox-LDL formation. Since we observed the accumulation and activation of neutrophils in HM-tumors, we evaluated the involvement of LOX-1 in neutrophil migration and activation. LOX-1 induced neutrophil migration via CCL2 secretion from TECs, which was enhanced by ox-LDL. Finally, we show genetic manipulation of LOX-1 expression in TECs or tumor stroma tended to reduce lung metastasis. Thus, the LOX-1/ox-LDL axis in TECs may lead to the formation of a high metastatic-tumor microenvironment via attracting neutrophils.
• Application of "Dredging Method" for the patients with odontogenic keratocyst.(和訳中)

  Ohiro Yoichi, Kakuguchi Wataru, Moritani Yasuhito, Kitamura Tetsuya, Maishi Nako, Hida Kyoko

  北海道歯学雑誌 43 63 - 69 0914-7063 2022/09
• The oral bacterium Streptococcus mutans promotes tumor metastasis by inducing vascular inflammation

  Li Yu, Nako Maishi, Erika Akahori, Akira Hasebe, Ryo Takeda, Aya Yanagawa Matsuda, Yasuhiro Hida, Jin‐Min Nam, Yasuhito Onodera, Yoshimasa Kitagawa, Kyoko Hida

  Cancer Science 1 (15) 1347-9032 2022/08/23 [Refereed][Not invited]
  
• Antiviral effect of cetylpyridinium chloride in mouthwash on SARS-CoV-2

  Ryo Takeda, Hirofumi Sawa, Michihito Sasaki, Yasuko Orba, Nako Maishi, Takuya Tsumita, Natsumi Ushijima, Yasuhiro Hida, Hidehiko Sano, Yoshimasa Kitagawa, Kyoko Hida

  Scientific Reports 12 (1) 14050 - 14050 2022/08/18 [Refereed]
   

  Abstract Cetylpyridinium chloride (CPC), a quaternary ammonium compound, which is present in mouthwash, is effective against bacteria, fungi, and enveloped viruses. This study was conducted to explore the antiviral effect of CPC on SARS-CoV-2. There are few reports on the effect of CPC against wild-type SARS-CoV-2 at low concentrations such as 0.001%–0.005% (10–50 µg/mL). Interestingly, we found that low concentrations of CPC suppressed the infectivity of human isolated SARS-CoV-2 strains (Wuhan, Alpha, Beta, and Gamma) even in saliva. Furthermore, we demonstrated that CPC shows anti-SARS-CoV-2 effects without disrupting the virus envelope, using sucrose density analysis and electron microscopic examination. In conclusion, this study provided experimental evidence that CPC may inhibit SARS-CoV-2 infection even at lower concentrations.
• 舌扁平上皮癌T1-2N0(UICC第8版)におけるリンパ節後発転移の有無に関する臨床病理学的検討

  格口 渉, 足利 雄一, 松田 彩, 栗林 和代, 間石 奈湖, 樋田 京子, 大廣 洋一

  頭頸部癌 (一社)日本頭頸部癌学会 48 (2) 215 - 215 1349-5747 2022/05
• Novel antiangiogenic therapy targeting biglycan using tumor endothelial cell‐specific liposomal siRNA delivery system

  Nako Maishi, Yu Sakurai, Hiroto Hatakeyama, Yui Umeyama, Takashi Nakamura, Rikito Endo, Mohammad Towfik Alam, Cong Li, Dorcas Akuba‐Muhyia Annan, Hiroshi Kikuchi, Hirofumi Morimoto, Masahiro Morimoto, Kosuke Akiyama, Noritaka Ohga, Yasuhiro Hida, Hideyoshi Harashima, Kyoko Hida

  Cancer Science 113 (5) 1855 - 1867 1347-9032 2022/05 [Refereed]
   

  Tumor blood vessels play important roles in tumor progression and metastasis. Targeting tumor endothelial cells (TECs) is one of the strategies for cancer therapy. We previously reported that biglycan, a small leucine-rich proteoglycan, is highly expressed in TECs. TECs utilize biglycan in an autocrine manner for migration and angiogenesis. Furthermore, TEC-derived biglycan stimulates tumor cell migration in a paracrine manner leading to tumor cell intravasation and metastasis. In this study, we explored the therapeutic effect of biglycan inhibition in the TECs of renal cell carcinoma using an in vivo siRNA delivery system known as a multifunctional envelope-type nanodevice (MEND), which contains a unique pH-sensitive cationic lipid. To specifically deliver MEND into TECs, we incorporated cyclo(Arg-Gly-Asp-D-Phe-Lys) (cRGD) into MEND because αV β3 integrin, a receptor for cRGD, is selective and highly expressed in TECs. We developed RGD-MEND-encapsulating siRNA against biglycan. First, we confirmed that MEND was delivered into OS-RC-2 tumor-derived TECs and induced in vitro RNAi-mediated gene silencing. MEND was then injected intravenously into OS-RC-2 tumor-bearing mice. Flow cytometry analysis demonstrated that MEND was specifically delivered into TECs. Quantitative RT-PCR indicated that biglycan was knocked down by biglycan siRNA-containing MEND. Finally, we analyzed the therapeutic effect of biglycan silencing by MEND in TECs. Tumor growth was inhibited by biglycan siRNA-containing MEND. Tumor microenvironmental factors such as fibrosis were also normalized using biglycan inhibition in TECs. Biglycan in TECs can be a novel target for cancer treatment.
• Editorial: Tumor Vessels as Directors of the Tumor Microenvironment: New Findings, Current Challenges & Perspectives

  Lucas Treps, Ann Ager, Kyoko Hida

  Frontiers in Cell and Developmental Biology 10 2022/03/29 [Refereed]
  
• miRNA-1246 in extracellular vesicles secreted from metastatic tumor induces drug resistance in tumor endothelial cells

  Chisaho Torii, Nako Maishi, Taisuke Kawamoto, Masahiro Morimoto, Kosuke Akiyama, Yusuke Yoshioka, Takashi Minami, Takuya Tsumita, Mohammad Towfik Alam, Takahiro Ochiya, Yasuhiro Hida, Kyoko Hida

  Scientific Reports 11 (1) 13502 - 13502 2021/12 [Refereed]
   

  Tumor endothelial cells (TECs) reportedly exhibit altered phenotypes. We have demonstrated that TECs acquire drug resistance with the upregulation of P-glycoprotein (P-gp, ABCB1), contrary to traditional assumptions. Furthermore, P-gp expression was higher in TECs of highly metastatic tumors than in those of low metastatic tumors. However, the detailed mechanism of differential P-gp expression in TECs remains unclear. miRNA was identified in highly metastatic tumor extracellular vesicles (EVs) and the roles of miRNA in endothelial cell resistance were analyzed in vitro and in vivo. In the present study, we found that treatment of highly metastatic tumor-conditioned medium induced resistance to 5-fluorouracil (5-FU) with interleukin-6 (IL-6) upregulation in endothelial cells (ECs). Among the soluble factors secreted from highly metastatic tumors, we focused on EVs and determined that miR-1246 was contained at a higher level in highly metastatic tumor EVs than in low metastatic tumor EVs. Furthermore, miR-1246 was transported via the EVs into ECs and induced IL-6 expression. Upregulated IL-6 induced resistance to 5-FU with STAT3 and Akt activation in ECs in an autocrine manner. These results suggested that highly metastatic tumors induce drug resistance in ECs by transporting miR-1246 through EVs.
• Inhibition of stromal biglycan promotes normalization of the tumor microenvironment and enhances chemotherapeutic efficacy

  Li Cong, Nako Maishi, Dorcas A. Annan, Marian F. Young, Hirofumi Morimoto, Masahiro Morimoto, Jin-Min Nam, Yasuhiro Hida, Kyoko Hida

  Breast Cancer Research 23 (1) 51 - 51 2021/12 [Refereed]
   

  Biglycan is a proteoglycan found in the extracellular matrix. We have previously shown that biglycan is secreted from tumor endothelial cells and induces tumor angiogenesis and metastasis. However, the function of stroma biglycan in breast cancer is still unclear. Biglycan gene analysis and its prognostic values in human breast cancers were based on TCGA data. E0771 breast cancer cells were injected into WT and Bgn KO mice, respectively. Breast cancer patients with high biglycan expression had worse distant metastasis-free survival. Furthermore, biglycan expression was higher in the tumor stromal compartment compared to the epithelial compartment. Knockout of biglycan in the stroma (Bgn KO) in E0771 tumor-bearing mice inhibited metastasis to the lung. Bgn KO also impaired tumor angiogenesis and normalized tumor vasculature by repressing tumor necrosis factor-ɑ/angiopoietin 2 signaling. Moreover, fibrosis was suppressed and CD8+ T cell infiltration was increased in tumor-bearing Bgn KO mice. Furthermore, chemotherapy drug delivery and efficacy were improved in vivo in Bgn KO mice. Our results suggest that targeting stromal biglycan may yield a potent and superior anticancer effect in breast cancer.
• Facile and rapid detection of SARS-CoV-2 antibody based on a noncompetitive fluorescence polarization immunoassay in human serum samples

  Keine Nishiyama, Kazuki Takahashi, Mao Fukuyama, Motohiro Kasuya, Ayuko Imai, Takumi Usukura, Nako Maishi, Masatoshi Maeki, Akihiko Ishida, Hirofumi Tani, Kyoko Hida, Koji Shigemura, Akihide Hibara, Manabu Tokeshi

  Biosensors and Bioelectronics 190 113414 - 113414 0956-5663 2021/10 [Refereed]
  
• Correlation between endothelial CXCR7 expression and clinicopathological factors in oral squamous cell carcinoma

  Misa Yanagiya, Randa I. H. Dawood, Nako Maishi, Yasuhiro Hida, Chisaho Torii, Dorcas A. Annan, Hiroshi Kikuchi, Aya Yanagawa Matsuda, Tetsuya Kitamura, Yoichi Ohiro, Masanobu Shindoh, Shinya Tanaka, Yoshimasa Kitagawa, Kyoko Hida

  Pathology International 71 (6) 383 - 391 1320-5463 2021/06 [Refereed]
   

  Oral squamous cell carcinoma (OSCC) impairs functionality and sensuousness resulting in poor quality of life. Biomarkers can predict disease trajectory and lead to effective treatments. Transcriptomics have identified genes that are upregulated in tumor endothelial cells (TECs) compared with normal endothelial cells (NECs). Among them, chemokine receptor 7 (CXCR7) is highly expressed in TECs of several cancers and involved in angiogenesis of TECs. However, levels of CXCR7 in OSCC blood vessels have not been fully investigated. In this study, we analyzed the correlation between CXCR7 expression in TECs and clinicopathological factors in OSCC. Immunohistochemistry for CXCR7 and CD34 was performed on 59 OSCC tissue specimens resected between 1996 and 2008 at Hokkaido University Hospital. CXCR7 expression in blood vessels was evaluated by the ratio of CXCR7+/CD34+ blood vessels. CXCR7 expression was 42% and 19% in tumor and non-tumor parts, respectively, suggesting that CXCR7 expression is higher in TECs than in NECs. CXCR7 expression in TECs correlated with advanced T-stage and cancer stage. Overall survival and disease-free survival rates were higher in low-expressing CXCR7 patients than in high-expressing. These results suggest that CXCR7 expression in blood vessels may be a useful diagnostic and prognostic marker for OSCC patients.
• The Role of IGF/IGF-IR-Signaling and Extracellular Matrix Effectors in Bone Sarcoma Pathogenesis

  George N. Tzanakakis, Eirini-Maria Giatagana, Aikaterini Berdiaki, Ioanna Spyridaki, Kyoko Hida, Monica Neagu, Aristidis M. Tsatsakis, Dragana Nikitovic

  Cancers 13 (10) 2478 - 2478 2021/05/19 [Refereed]
   

  Bone sarcomas, mesenchymal origin tumors, represent a substantial group of varying neoplasms of a distinct entity. Bone sarcoma patients show a limited response or do not respond to chemotherapy. Notably, developing efficient chemotherapy approaches, dealing with chemoresistance, and preventing metastasis pose unmet challenges in sarcoma therapy. Insulin-like growth factors 1 and 2 (IGF-1 and -2) and their respective receptors are a multifactorial system that significantly contributes to bone sarcoma pathogenesis. Whereas failures have been registered in creating novel targeted therapeutics aiming at the IGF pathway, new agent development should continue, evaluating combinatorial strategies for enhancing antitumor responses and better classifying the patients that could best benefit from these therapies. A plausible approach for developing a combinatorial strategy is to focus on the tumor microenvironment (TME) and processes executed therein. Herewith, we will discuss how the interplay between IGF-signaling and the TME constituents affects sarcomas’ basal functions and their response to therapy. This review highlights key studies focusing on IGF signaling in bone sarcomas, specifically studies underscoring novel properties that make this system an attractive therapeutic target and identifies new relationships that may be exploited. Potential direct and adjunct therapeutical implications of the extracellular matrix (ECM) effectors will also be summarized.
• A novel dual-targeted rosiglitazone-loaded nanoparticle for the prevention of diet-induced obesity via the browning of white adipose tissue

  Ryu Hiradate, Ikramy A. Khalil, Aya Matsuda, Mika Sasaki, Kyoko Hida, Hideyoshi Harashima

  Journal of Controlled Release 329 665 - 675 0168-3659 2021/01
• Biglycan, tumor endothelial cell secreting proteoglycan, as possible biomarker for lung cancer,

  Morimoto H, Hida Y, Maishi N, Nishihara H, Hatanaka Y, Li C, Matsuno Y, Nakamura T, Hirano S, Hida K

  Thoracic cancer 12 (9) 1347 - 1357 2021 [Refereed]
   

  OBJECTIVES: In lung cancer, surgery remains the most curative treatment and limited resection is beneficial for patients with low cardiopulmonary function and low malignancy tumors. However, there are no biomarkers of low malignancy to select candidates for limited resection without compromising the outcome of treatments. Recently we identified biglycan (BGN) as a tumor endothelial cell (TEC) marker that is associated with tumor progression in various cancers. In this study, we analyzed the association between BGN expression in TECs in lung cancer and cancer progression in patients. MATERIALS AND METHODS: First, we performed immunohistochemistry of BGN with resected lung tumor tissues of 155 patients who had undergone thoracic surgery and analyzed the correlation between BGN-positive vessel density in primary lung tumors and clinicopathological factors. Second, we measured the BGN levels in preoperative serum of other 46 patients with lung cancer by ELISA, and analyzed the correlation between BGN expression in tumor tissues and blood BGN levels. RESULTS: High BGN expression in the TECs was significantly associated with T factor, and was a significant negative predictor. BGN levels in preoperative serum of 46 patients with lung cancer was significantly correlated with BGN expression in the TECs. Preoperative serum BGN level was significantly lower in healthy volunteers and less invasive adenocarcinoma than in invasive adenocarcinoma and other lung carcinomas. These results suggest that low BGN level in preoperative serum in patients with lung cancer might indicate low malignancy. CONCLUSIONS: BGN can be a potential biomarker for lung cancer.
• グルタミンの代謝はレドックスホメオスタシスを介して腫瘍内皮細胞の増殖を促進する(Glutamine metabolism facilitates tumor endothelial cell proliferation via redox homeostasis)

  Annan Dorcas A., 間石 奈湖, 曽我 朋義, Hojo Takayuki, 樋田 泰浩, 樋田 京子

  日本癌学会総会記事 79回 OE10 - 2 0546-0476 2020/10
• 肺癌マウスモデルにおけるBiglycanの腫瘍微小環境に及ぼす影響

  佐藤 峰嘉, Li Cong, 間石 奈湖, Annan Dorcas A., 樋田 泰浩, 樋田 京子

  日本癌学会総会記事 79回 OE10 - 5 0546-0476 2020/10
• 腫瘍血管内皮細胞が発現するLOX-1のがん転移における役割

  積田 卓也, Annan Dorcas A., 間石 奈湖, 樋田 泰浩, 樋田 京子

  日本癌学会総会記事 79回 OJ10 - 7 0546-0476 2020/10
• リポソームsiRNAデリバリーシステムを用いた腫瘍血管biglycanを標的とした新規血管新生阻害療法

  間石 奈湖, 櫻井 遊, 畠山 浩人, Li Cong, Alam Mohammad T., 菊地 央, 森本 浩史, 森本 真弘, 樋田 泰浩, 原島 秀吉, 樋田 京子

  日本癌学会総会記事 79回 OE16 - 5 0546-0476 2020/10
• 膵癌の腫瘍血管内皮細胞におけるbiglycan発現の臨床的意義の検討

  田中 宏典, 間石 奈湖, 森本 浩史, Annan Dorcas A., 中村 透, 樋田 泰浩, 平野 聡, 樋田 京子

  日本癌学会総会記事 79回 PJ14 - 10 0546-0476 2020/10
• Chemotherapy-Induced IL8 Upregulates MDR1/ABCB1 in Tumor Blood Vessels and Results in Unfavorable Outcome.

  Hiroshi Kikuchi, Nako Maishi, Dorcas A Annan, Mohammad Towfik Alam, Randa Ibrahim Hassan Dawood, Masumi Sato, Masahiro Morimoto, Ryo Takeda, Keita Ishizuka, Ryuji Matsumoto, Tomoshige Akino, Kunihiko Tsuchiya, Takashige Abe, Takahiro Osawa, Naoto Miyajima, Satoru Maruyama, Toru Harabayashi, Manabu Azuma, Katsushige Yamashiro, Kaname Ameda, Akira Kashiwagi, Yoshihiro Matsuno, Yasuhiro Hida, Nobuo Shinohara, Kyoko Hida

  Cancer research 80 (14) 2996 - 3008 0008-5472 2020/07/15 [Refereed][Not invited]
   

  Tumor endothelial cells (TEC) lining tumor blood vessels actively contribute to tumor progression and metastasis. In addition to tumor cells, TEC may develop drug resistance during cancer treatment, allowing the tumor cells to survive chemotherapy and metastasize. We previously reported that TECs resist paclitaxel treatment via upregulation of ABCB1. However, whether TEC phenotypes are altered by anticancer drugs remains to be clarified. Here, we show that ABCB1 expression increases after chemotherapy in urothelial carcinoma cases. The ratio of ABCB1-positive TEC before and after first-line chemotherapy in urothelial carcinoma tissues (n = 66) was analyzed by ABCB1 and CD31 immunostaining. In 42 cases (64%), this ratio increased after first-line chemotherapy. Chemotherapy elevated ABCB1 expression in endothelial cells by increasing tumor IL8 secretion. In clinical cases, ABCB1 expression in TEC correlated with IL8 expression in tumor cells after first-line chemotherapy, leading to poor prognosis. In vivo, the ABCB1 inhibitor combined with paclitaxel reduced tumor growth and metastasis compared with paclitaxel alone. Chemotherapy is suggested to cause inflammatory changes in tumors, inducing ABCB1 expression in TEC and conferring drug resistance. Overall, these findings indicate that TEC can survive during chemotherapy and provide a gateway for cancer metastasis. Targeting ABCB1 in TEC represents a novel strategy to overcome cancer drug resistance. SIGNIFICANCE: These findings show that inhibition of ABCB1 in tumor endothelial cells may improve clinical outcome, where ABCB1 expression contributes to drug resistance and metastasis following first-line chemotherapy.
• Tumor Endothelial Cell-A Biological Tool for Translational Cancer Research.

  Dorcas Akuba-Muhyia Annan, Hiroshi Kikuchi, Nako Maishi, Yasuhiro Hida, Kyoko Hida

  International journal of molecular sciences 21 (9) 1661-6596 2020/05/03 [Refereed][Not invited]
   

  Going from bench to bedside is a simplified description of translational research, with the ultimate goal being to improve the health status of mankind. Tumor endothelial cells (TECs) perform angiogenesis to support the growth, establishment, and dissemination of tumors to distant organs. TECs have various features that distinguish them from normal endothelial cells, which include alterations in gene expression patterns, higher angiogenic and metabolic activities, and drug resistance tendencies. The special characteristics of TECs enhance the vulnerability of tumor blood vessels toward antiangiogenic therapeutic strategies. Therefore, apart from being a viable therapeutic target, TECs would act as a better mediator between the bench (i.e., angiogenesis research) and the bedside (i.e., clinical application of drugs discovered through research). Exploitation of TEC characteristics could reveal unidentified strategies of enhancing and monitoring antiangiogenic therapy in the treatment of cancer, which are discussed in this review.
• Tumor blood vessels as targets for cancer therapy

  Kyoko Hida, Nako Maishi, Yasuhiro Hida

  Cancer Drug Delivery Systems Based on the Tumor Microenvironment 41 - 56 2020/01/01 [Refereed][Not invited]
   

  © Springer Japan KK, part of Springer Nature 2019. Tumor growth and metastasis are dependent on angiogenesis, which is the formation of new blood vessels. The newly formed blood vessels around the tumor supply oxygen and nutrients to the tumor, supporting its progression. Moreover, these blood vessels also serve as channels through which tumor cells metastasize to distant organs. Tumor blood vessels, and especially the endothelial cells lining tumor blood vessels [tumor endothelial cells (TECs)], are important targets in cancer therapy. Since newly formed tumor blood vessels originate from pre-existing normal vessels, tumor blood vessels and TECs traditionally have been considered the same as normal vasculature. Tumor blood vessels, however, have a distinctively abnormal phenotype, including morphological alterations. Recently, it has been revealed that TECs constitute a heterogeneous population, exhibiting characteristics induced largely by tumor microenvironmental factors. Furthermore, TECs induce cancer progression through metastasis passively but also actively. This chapter will review the mechanisms underlying tumor angiogenesis and discuss the biology of TECs, offering new perspectives on treatment strategies that can target tumor blood vessels from a personalized medicine perspective.
• TNF-α enhances TGF-β-induced endothelial-to-mesenchymal transition via TGF-β signal augmentation

  Yasuhiro Yoshimatsu, Ikumi Wakabayashi, Shiori Kimuro, Naoya Takahashi, Kazuki Takahashi, Miho Kobayashi, Nako Maishi, Katarzyna A. Podyma‐Inoue, Kyoko Hida, Kohei Miyazono, Tetsuro Watabe

  Cancer Science 111 (7) 2385 - 2399 1347-9032 2020 [Refereed][Not invited]
  
• Role of Dimerized C16orf74 in Aggressive Pancreatic Cancer: A Novel Therapeutic Target

  Toshihiro Kushibiki, Toru Nakamura, Masumi Tsuda, Takahiro Tsuchikawa, Koji Hontani, Kazuho Inoko, Mizuna Takahashi, Toshimichi Asano, Keisuke Okamura, Soichi Murakami, Yo Kurashima, Yuma Ebihara, Takehiro Noji, Yoshitsugu Nakanishi, Kimitaka Tanaka, Nako Maishi, Katsunori Sasaki, Woong-Ryeon Park, Toshiaki Shichinohe, Kyoko Hida, Shinya Tanaka, Satoshi Hirano

  Molecular Cancer Therapeutics 19 (1) 187 - 198 1535-7163 2020/01/01 [Refereed]
   

  Abstract Over the past 30 years, the therapeutic outcome for pancreatic ductal adenocarcinoma (PDAC) has remained stagnant due to the lack of effective treatments. We performed a genome-wide analysis to identify novel therapeutic targets for PDAC. Our analysis showed that Homo sapiens chromosome 16 open reading frame 74 (C16orf74) was upregulated in most patients with PDAC and associated with poor prognosis. Previously, we demonstrated that C16orf74 interacts with the catalytic subunit alpha of protein phosphatase 3 and plays an important role in PDAC invasion. However, the pathophysiologic function of C16orf74 is still unclear. In this study, through the analysis of C16orf74 interaction, we demonstrate a new strategy to inhibit the growth and invasion of PDAC. C16orf74 exists in the homodimer form under the cell membrane and binds integrin αVβ3 and is also involved in invasion by activating Rho family (Rac1) and MMP2. Considering that this dimeric form was found to be involved in the function of C16orf74, we designed an 11R-DB (dimer block) cell-permeable dominant-negative peptide that inhibits the dimer form of C16orf74. 11R-DB suppressed invasion and proliferation of PDAC cell lines by inhibiting phosphorylation of Akt and mTOR and also by inactivation of MMP2. 11R-DB also showed antitumor effects in an orthotopic xenograft model and peritoneal metastasis model. Thus, this study demonstrates that dimerized C16orf74, present in the cell membrane, is involved in pancreatic cancer invasion and proliferation. In addition, the C16orf74 dimer block cell-permeable peptide (11R-DB) has a potent therapeutic effect on PDAC in vitro and in vivo.
• Carbonic anhydrase 2 (CAII) supports tumor blood endothelial cell survival under lactic acidosis in the tumor microenvironment.

  Dorcas A Annan, Nako Maishi, Tomoyoshi Soga, Randa Dawood, Cong Li, Hiroshi Kikuchi, Takayuki Hojo, Masahiro Morimoto, Tetsuya Kitamura, Mohammad Towfik Alam, Kazuyuki Minowa, Nobuo Shinohara, Jin-Min Nam, Yasuhiro Hida, Kyoko Hida

  Cell communication and signaling : CCS 17 (1) 169 - 169 2019/12/17 [Refereed][Not invited]
   

  BACKGROUND: Tumor endothelial cells (TECs) perform tumor angiogenesis, which is essential for tumor growth and metastasis. Tumor cells produce large amounts of lactic acid from glycolysis; however, the mechanism underlying the survival of TECs to enable tumor angiogenesis under high lactic acid conditions in tumors remains poorly understood. METHODOLOGY: The metabolomes of TECs and normal endothelial cells (NECs) were analyzed by capillary electrophoresis time-of-flight mass spectrometry. The expressions of pH regulators in TECs and NECs were determined by quantitative reverse transcription-PCR. Cell proliferation was measured by the MTS assay. Western blotting and ELISA were used to validate monocarboxylate transporter 1 and carbonic anhydrase 2 (CAII) protein expression within the cells, respectively. Human tumor xenograft models were used to access the effect of CA inhibition on tumor angiogenesis. Immunohistochemical staining was used to observe CAII expression, quantify tumor microvasculature, microvessel pericyte coverage, and hypoxia. RESULTS: The present study shows that, unlike NECs, TECs proliferate in lactic acidic. TECs showed an upregulated CAII expression both in vitro and in vivo. CAII knockdown decreased TEC survival under lactic acidosis and nutrient-replete conditions. Vascular endothelial growth factor A and vascular endothelial growth factor receptor signaling induced CAII expression in NECs. CAII inhibition with acetazolamide minimally reduced tumor angiogenesis in vivo. However, matured blood vessel number increased after acetazolamide treatment, similar to bevacizumab treatment. Additionally, acetazolamide-treated mice showed decreased lung metastasis. CONCLUSION: These findings suggest that due to their effect on blood vessel maturity, pH regulators like CAII are promising targets of antiangiogenic therapy. Video Abstract.
• Development of Immortalized Human Tumor Endothelial Cells from Renal Cancer.

  Nako Maishi, Hiroshi Kikuchi, Masumi Sato, Hiroko Nagao-Kitamoto, Dorcas A Annan, Shogo Baba, Takayuki Hojo, Misa Yanagiya, Yusuke Ohba, Genichiro Ishii, Kenkichi Masutomi, Nobuo Shinohara, Yasuhiro Hida, Kyoko Hida

  International journal of molecular sciences 20 (18) 4595  2019/09/17 [Refereed][Not invited]
   

  Tumor angiogenesis research and antiangiogenic drug development make use of cultured endothelial cells (ECs) including the human microvascular ECs among others. However, it has been reported that tumor ECs (TECs) are different from normal ECs (NECs). To functionally validate antiangiogenic drugs, cultured TECs are indispensable tools, but are not commercially available. Primary human TECs are available only in small quantities from surgical specimens and have a short life span in vitro due to their cellular senescence. We established immortalized human TECs (h-imTECs) and their normal counterparts (h-imNECs) by infection with lentivirus producing simian virus 40 large T antigen and human telomerase reverse transcriptase to overcome the replication barriers. These ECs exhibited an extended life span and retained their characteristic endothelial morphology, expression of endothelial marker, and ability of tube formation. Furthermore, h-imTECs showed their specific characteristics as TECs, such as increased proliferation and upregulation of TEC markers. Treatment with bevacizumab, an antiangiogenic drug, dramatically decreased h-imTEC survival, whereas the same treatment failed to alter immortalized NEC survival. Hence, these h-imTECs could be a valuable tool for drug screening to develop novel therapeutic agents specific to TECs or functional biological assays in tumor angiogenesis research.
• Fibroblast growth factor signals regulate transforming growth factor-β-induced endothelial-to-myofibroblast transition of tumor endothelial cells via Elk1.

  Akatsu Y, Takahashi N, Yoshimatsu Y, Kimuro S, Muramatsu T, Katsura A, Maishi N, Suzuki HI, Inazawa J, Hida K, Miyazono K, Watabe T

  Mol Oncol. 13 (8) 1706 - 1724 1574-7891 2019/08 [Refereed][Not invited]
  
• Two modes of toxicity of lipid nanoparticles containing a pH-sensitive cationic lipid on human A375 and A375-SM melanoma cell lines

  AlBaloul A, Sato Y, Maishi N, Hida K, Harashima H

  BPB Reports 2 48 - 55 2019 [Refereed][Not invited]
  
• Contribution of tumor endothelial cells in cancer progression

  Kyoko Hida, Nako Maishi, Dorcas A. Annan, Yasuhiro Hida

  International Journal of Molecular Sciences 19 (5) 1272  1661-6596 2018/05 [Refereed][Not invited]
   

  © 2018 by the authors. Licensee MDPI, Basel, Switzerland. Tumor progression depends on the process of angiogenesis, which is the formation of new blood vessels. These newly formed blood vessels supply oxygen and nutrients to the tumor, supporting its progression and providing a gateway for tumor metastasis. Tumor angiogenesis is regulated by the balance between angiogenic activators and inhibitors within the tumor microenvironment. Because the newly formed tumor blood vessels originate from preexisting normal vessels, tumor blood vessels, and tumor endothelial cells (TECs) have historically been considered to be the same as normal blood vessels and endothelial cells; however, evidence of TECs’ distinctive abnormal phenotypes has increased. In addition, it has been revealed that TECs constitute a heterogeneous population. Thus, TECs that line tumor blood vessels are important targets in cancer therapy. We have previously reported that TECs induce cancer metastasis. In this review, we describe recent studies on TEC abnormalities related to cancer progression to provide insight into new anticancer therapies.
• Tumor endothelial cells with high aldehyde dehydrogenase activity show drug resistance.

  Kyoko Hida, Nako Maishi, Kosuke Akiyama, Hitomi Ohmura-Kakutani, Chisaho Torii, Noritaka Ohga, Takahiro Osawa, Hiroshi Kikuchi, Hirofumi Morimoto, Masahiro Morimoto, Masanobu Shindoh, Nobuo Shinohara, Yasuhiro Hida

  Cancer science 108 (11) 2195 - 2203 1347-9032 2017/11 [Refereed][Not invited]
   

  Tumor blood vessels play an important role in tumor progression and metastasis. We previously reported that tumor endothelial cells (TEC) exhibit several altered phenotypes compared with normal endothelial cells (NEC). For example, TEC have chromosomal abnormalities and are resistant to several anticancer drugs. Furthermore, TEC contain stem cell-like populations with high aldehyde dehydrogenase (ALDH) activity (ALDHhigh TEC). ALDHhigh TEC have proangiogenic properties compared with ALDHlow TEC. However, the association between ALDHhigh TEC and drug resistance remains unclear. In the present study, we found that ALDH mRNA expression and activity were higher in both human and mouse TEC than in NEC. Human NEC:human microvascular endothelial cells (HMVEC) were treated with tumor-conditioned medium (tumor CM). The ALDHhigh population increased along with upregulation of stem-related genes such as multidrug resistance 1, CD90, ALP, and Oct-4. Tumor CM also induced sphere-forming ability in HMVEC. Platelet-derived growth factor (PDGF)-A in tumor CM was shown to induce ALDH expression in HMVEC. Finally, ALDHhigh TEC were resistant to fluorouracil (5-FU) in vitro and in vivo. ALDHhigh TEC showed a higher grade of aneuploidy compared with that in ALDHlow TEC. These results suggested that tumor-secreting factor increases ALDHhigh TEC populations that are resistant to 5-FU. Therefore, ALDHhigh TEC in tumor blood vessels might be an important target to overcome or prevent drug resistance.
• Abnormalities of tumor endothelial cells and cancer progression

  Hida K, Maishi N

  Oral Sci Int 15 (1) 1 - 6 2017/10 [Refereed][Not invited]
  
• Tumor endothelial cells accelerate tumor metastasis

  Nako Maishi, Kyoko Hida

  CANCER SCIENCE 108 (10) 1921 - 1926 1349-7006 2017/10 [Refereed][Not invited]
   

  Tumor metastasis is the main cause of cancer-related death. Understanding the molecular mechanisms underlying tumor metastasis is crucial to control this fatal disease. Several molecular pathways orchestrate the complex biological cell events during a metastatic cascade. It is now well known that bidirectional interaction between tumor cells and their microenvironment, including tumor stroma, is important for tumor progression and metastasis. Tumor stromal cells, which acquire their specific characteristics in the tumor microenvironment, accelerate tumor malignancy. The formation of new blood vessels, termed as tumor angiogenesis, is a requirement for tumor progression. Tumor blood vessels supply nutrients and oxygen and also provide the route for metastasis. Tumor endothelial cells, which line tumor blood vessels, also exhibit several altered phenotypes compared with those of their normal counterparts. Recent studies have emphasized angiocrine factors that are released from tumor endothelial cells and promote tumor progression. During intravasation, tumor cells physically contact tumor endothelial cells and interact with them by juxtacrine and paracrine signaling. Recently, we observed that in highly metastatic tumors, tumor endothelial cells interact with tumor cells by secretion of a small leucine-rich repeat proteoglycan known as biglycan. Biglycan from tumor endothelial cells stimulates the tumor cells to metastasize. In the present review, we highlight the role of tumor stromal cells, particularly endothelial cells, in the initial steps of tumor metastasis.
• ATP-binding cassette transporters in tumor endothelial cells and resistance to metronomic chemotherapy

  Kyoko Hida, Hiroshi Kikuchi, Nako Maishi, Yasuhiro Hida

  CANCER LETTERS 400 305 - 310 0304-3835 2017/08 [Refereed][Invited]
   

  Drug resistance is a major problem in anticancer therapy. ATP-binding cassette (ABC) transporters have a role in the multidrug resistance. A new regimen of chemotherapy has been proposed, called "metronomic chemotherapy". Metronomic chemotherapy is the frequent, regular administration of drug doses designed to maintain low, but active, concentrations of chemotherapeutic drugs over prolonged periods of time, without causing serious toxicities. Metronomic chemotherapy regimens were developed to optimize the antitumor efficacy of agents that target the tumor vasculature instead of tumor cells, and to reduce toxicity of antineoplastic drugs [1]. Nevertheless, recent studies revealed that ABC transporters are expressed at a higher level in the endothelium in the tumor. To avoid resistance to metronomic antiangiogenic chemotherapy, ABC transporter inhibition of tumor endothelial cells may be a promising strategy. In this mini-review, we discuss the possible mechanism of resistance to metronomic chemotherapy from the viewpoint of tumor endothelial cell biology, focusing on ABC transporters. (C) 2017 Published by Elsevier Ireland Ltd.
• miR-145 promoted anoikis resistance in tumor endothelial cells

  Kyoko Hida, Taisuke Kawamoto, Nako Maishi, Masahiro Morimoto, Kosuke Akiyama, Noritaka Ohga, Masanobu Shindoh, Nobuo Shinohara, Yasuhiro Hida

  JOURNAL OF BIOCHEMISTRY 162 (2) 81 - 84 0021-924X 2017/08 [Refereed][Not invited]
   

  Tumor progression is dependent on tumor angiogenesis. We previously reported that the phenotype of tumor endothelial cells (TECs) is distinct from normal endothelial cells (NECs). Herein, we conducted a pathway analysis using a public TEC microarray database and identified several putative TEC-specific miRNAs. We found that miR-145 expression was upregulated in TECs and that miR-145 enhanced cell adhesion and anoikis resistance and upregulated Bcl-2 and Bcl-xl via ERK1/2 in human microvascular endothelial cells. These findings suggested that miR-145 is involved in the acquisition of the TEC phenotype, partially. Therefore, miR-145 and its target genes may be molecular targets for anti-angiogenic therapy.
• Vasohibin-1 as a Novel Prognostic Factor for Head and Neck Squamous Cell Carcinoma

  Chisaho Torii, Yasuhiro Hida, Masanobu Shindoh, Kosuke Akiyama, Noritaka Ohga, Nako Maishi, Yoichi Ohiro, Mitsunobu Ono, Yasunori Totsuka, Yoshimasa Kitagawa, Kanchu Tei, Yasufumi Sato, Kyoko Hida

  ANTICANCER RESEARCH 37 (3) 1219 - 1225 0250-7005 2017/03 [Refereed][Not invited]
   

  Aim: We evaluated the prognostic value of vasohibin-1 (VASH1) expression in head and neck squamous cell carcinoma. Materials and Methods: Immunohistochemistry for VASH1 and cluster of differentiation 34 (CD34) was performed on 61 head and neck squamous cell carcinoma specimens. The association between VASH1 expression in the tumour and clinical outcomes was analyzed statistically. Results: VASH1 staining in normal tissue adjacent to cancerous tissue was negative, whereas it was positive in tumour blood vessels and AE1/AE3 and Ki67-positive tumour cells. Therefore, we examined the association between VASH1 expression in the tumour and clinical outcomes. Patients with high VASH1 expression in tumour had significantly shorter disease-free survival and more frequently had lymph node recurrence than those with low VASH1 expression. Conclusion: These results suggest that VASH1 expression is associated with tumour progression and may be useful as a prognostic marker of head and neck squamous cell carcinoma.
• Aneuploidy of a murine immortalized endothelial cell line, MS1

  Kyoko Hida, Nako Maishi, Dorcas Akuba Muhyia Annan, Miyako Kondoh, Takayuki Hojo, Umma Habiba, Noritaka Ohga, Kosuke Ishikawa, Masumi Sato, Chisaho Torii, Misa Yanagiya, Masahiro Morimoto, Yasuhiro Hida, Masanobu Shindoh

  Journal of Oral Biosciences 59 (1) 50 - 54 1349-0079 2017/02/01 [Refereed][Not invited]
   

  © 2016 Japanese Association for Oral Biology Objectives We have previously reported that tumor endothelial cells (TECs) are chromosomally abnormal. It is generally known that immortalized cells have chromosomal abnormalities. In this study, our objective was to compare chromosomal properties of an immortalized normal endothelial cell (NEC) line, MS1, and those of TECs, to assess the possible usefulness of MS1 cells as a surrogate for TECs, which have some experimental intractability as primary cultured endothelial cells. Methods Primary cultured NECs were isolated from murine dermis tissue. Fluorescence in situ hybridization was performed to analyze aneuploidy in MS1 cells and NECs. DNA damage response (DDR) activation was analyzed by γH2AX staining. We also tested the involvement of reactive oxygen species (ROS) in the chromosomal abnormalities of MS1 cells. Results MS1 cells showed a higher rate of aneuploidy than NECs did (91.6% versus 7.6%). A DDR was activated to a greater extent in MS1 cells than in NECs, judging by H2AX phosphorylation. ROS induced H2AX activation in MS1 cells, suggesting that ROS are involved in their DDR. Conclusion MS1 cells are aneuploid and activate H2AX to a greater extent than NECs do. MS1 cells resemble TECs in terms of the proliferative phenotype and aneuploidy. Thus, MS1 may be a good cell line for studies on the relation between chromosomal instability and a DDR in nonmalignant cells.
• ROS enhance angiogenic properties via regulation of NRF2 in tumor endothelial cells

  Takayuki Hojo, Nako Maishi, Alam Mohammad Towfik, Kosuke Akiyama, Noritaka Ohga, Masanobu Shindoh, Yasuhiro Hida, Kazuyuki Minowa, Toshiaki Fujisawa, Kyoko Hida

  Oncotarget 8 (28) 45484 - 45495 2017 [Refereed][Not invited]
   

  © Hojo et al. Reactive oxygen species (ROS) are unstable molecules that activate oxidative stress. Because of the insufficient blood flow in tumors, the tumor microenvironment is often exposed to hypoxic condition and nutrient deprivation, which induces ROS accumulation. We isolated tumor endothelial cells (TECs) and found that they have various abnormalities, although the underlying mechanisms are not fully understood. Here we showed that ROS were accumulated in tumor blood vessels and ROS enhanced TEC migration with upregulation of several angiogenesis related gene expressions. It was also demonstrated that these genes were upregulated by regulation of Nuclear factor erythroid 2-related factor 2 (NRF2). Among these genes, we focused on Biglycan, a small leucine-rich proteoglycan. Inhibition of Toll-like receptors 2 and 4, known BIGLYCAN (BGN) receptors, cancelled the TEC motility stimulated by ROS. ROS inhibited NRF2 expression in TECs but not in NECs, and NRF2 inhibited phosphorylation of SMAD2/3, which activates transcription of BGN. These results indicated that ROS-induced BGN caused the pro-angiogenic phenotype in TECs via NRF2 dysregulation.
• ALDH1 and podoplanin expression patterns predict the risk of malignant transformation in oral leukoplakia

  Umma Habiba, Kyoko Hida, Tetsuya Kitamura, Aya Yanagawa Matsuda, Fumihiro Higashino, Yoichi M. Ito, Yoichi Ohiro, Yasunori Totsuka, Masanobu Shindoh

  ONCOLOGY LETTERS 13 (1) 321 - 328 1792-1074 2017/01 [Refereed][Not invited]
   

  Oral leukoplakia (OL) is a clinically diagnosed preneoplastic lesion of the oral cavity with an increased oral cancer risk. However, the risk of malignant transformation is still difficult to assess. The objective of the present study was to examine the expression patterns of aldehyde dehydrogenase 1 (ALDH1) and podoplanin in OL, and to determine their roles in predicting oral cancer development. In the present study, the expression patterns of ALDH1 and podoplanin were determined in samples from 79 patients with OL. The association between protein expression and clinicopathological parameters, including oral cancer-free survival, was analyzed during a mean follow-up period of 3.4 years. Expression of ALDH1 and podoplanin was observed in 61 and 67% patients, respectively. Kaplan-Meier analysis demonstrated that the expression of the proteins was correlated with the risk of progression to oral cancer. Multivariate analysis revealed that expression of ALDH1 and podoplanin was associated with 3.02- and 2.62-fold increased risk of malignant transformation, respectively. The malignant transformation risk of OL was considerably higher in cases with expression of both proteins. Point-prevalence analysis revealed that 66% of patients with co-expression of ALDH1 and podoplanin developed oral cancer. Taken together, our data indicate that ALDH1 and podoplanin expression patterns in OL are associated with oral cancer development, suggesting that ALDH1 and podoplanin may be useful biomarkers to identify OL patients with a substantially high oral cancer risk.
• Tumor endothelial cells express high pentraxin 3 levels

  Kyoko Hida, Nako Maishi, Taisuke Kawamoto, Kosuke Akiyama, Noritaka Ohga, Yasuhiro Hida, Kenji Yamada, Takayuki Hojo, Hiroshi Kikuchi, Masumi Sato, Chisaho Torii, Nobuo Shinohara, Masanobu Shindoh

  PATHOLOGY INTERNATIONAL 66 (12) 687 - 694 1320-5463 2016/12 [Refereed][Not invited]
   

  It has been described that tumor progression has many similarities to inflammation and wound healing in terms of the signaling processes involved. Among biological responses, angiogenesis, which is necessary for tumor progression and metastasis, is a common hallmark; therefore, tumor blood vessels have been considered as important therapeutic targets in anticancer therapy. We focused on pentraxin 3 (PTX3), which is a marker of cancer-related inflammation, but we found no reports on its expression and function in tumor blood vessels. Here we showed that PTX3 is expressed inmouse and human tumor blood vessels based on immunohistochemical analysis. We found that PTX3 is upregulated in primary mouse and human tumor endothelial cells compared to normal endothelial cells. We also showed that PTX3 plays an important role in the proliferation of the tumor endothelial cells. These results suggest that PTX3 is an important target for antiangiogenic therapy.
• HuR and podoplanin expression is associated with a high risk of malignant transformation in patients with oral preneoplastic lesions

  Umma Habiba, Tetsuya Kitamura, Aya Yanagawa-Matsuda, Fumihiro Higashino, Kyoko Hida, Yasunori Totsuka, Masanobu Shindoh

  ONCOLOGY LETTERS 12 (5) 3199 - 3207 1792-1074 2016/11 [Refereed][Not invited]
   

  The risk of malignant transformation in oral preneoplastic lesions (OPLs) is challenging to assess. The objective of the present study was to determine the expression of ELAV like RNA binding protein 1 (HuR) and podoplanin in OPLs, and to evaluate the use of each protein as biomarkers for the risk assessment of malignant transformations. Immunohistochemistry for HuR and podoplanin was performed on the tissues of 51 patients with OPL, including cases of low grade dysplasia (LGD) and high grade dysplasia (HGD). The association between the protein expression patterns and clinicopathological parameters, including oral cancer free survival (OCFS) time, was analyzed during the follow-up period. HuR and podoplanin expression was observed in 28 (55%) and 36 (71%) of 51 patients, respectively. Kaplan-Meier analysis showed that the expression of HuR and podoplanin was associated with the risk of progression to oral cancer (P<0.05). Multivariate analysis revealed that HuR and podoplanin expression was associated with a 2.93-fold (95% confidence interval (CI), 0.98-10.34; P=0.055) and 2.06-fold (95% CI, 0.55-8.01; P=0.283) increase in risk of malignant transformation, respectively. The risk of OPL malignant transformation was considerably increased with the coexpression of HuR and podoplanin compared with the histological grading (95% CI, 1.64-23.59; P=0.005). The results of the present study demonstrated that the expression of HuR and podoplanin associates with malignant transformation and suggests that the proteins may be used as biomarkers to identify OPL patients with an increased risk of cancer development.
• Tumour endothelial cells in high metastatic tumours promote metastasis via epigenetic dysregulation of biglycan

  Nako Maishi, Yusuke Ohba, Kosuke Akiyama, Noritaka Ohga, Jun-ichi Hamada, Hiroko Nagao-Kitamoto, Mohammad Towfik Alam, Kazuyuki Yamamoto, Taisuke Kawamoto, Nobuo Inoue, Akinobu Taketomi, Masanobu Shindoh, Yasuhiro Hida, Kyoko Hida

  SCIENTIFIC REPORTS 6 28039  2045-2322 2016/06 [Refereed][Not invited]
   

  Tumour blood vessels are gateways for distant metastasis. Recent studies have revealed that tumour endothelial cells (TECs) demonstrate distinct phenotypes from their normal counterparts. We have demonstrated that features of TECs are different depending on tumour malignancy, suggesting that TECs communicate with surrounding tumour cells. However, the contribution of TECs to metastasis has not been elucidated. Here, we show that TECs actively promote tumour metastasis through a bidirectional interaction between tumour cells and TECs. Co-implantation of TECs isolated from highly metastatic tumours accelerated lung metastases of low metastatic tumours. Biglycan, a small leucine-rich repeat proteoglycan secreted from TECs, activated tumour cell migration via nuclear factor-kappa B and extracellular signal-regulated kinase 1/2. Biglycan expression was upregulated by DNA demethylation in TECs. Collectively, our results demonstrate that TECs are altered in their microenvironment and, in turn, instigate tumour cells to metastasize, which is a novel mechanism for tumour metastasis.
• Tumor angiogenesis-characteristics of tumor endothelial cells

  Kyoko Hida, Nako Maishi, Chisaho Torii, Yasuhiro Hida

  INTERNATIONAL JOURNAL OF CLINICAL ONCOLOGY 21 (2) 206 - 212 1341-9625 2016/04 [Refereed][Not invited]
   

  Tumor blood vessels provide nutrition and oxygen to the tumor, resulting in tumor progression. They also act as gatekeepers, inducing tumor metastasis. Thus, targeting tumor blood vessels is an important strategy in cancer therapy. Tumor endothelial cells (TECs), which line the inner layer of blood vessels of the tumor stromal tissue, are the main targets of anti-angiogenic therapy. Because new tumor blood vessels generally sprout from pre-existing vasculature, they have been considered to be the same as normal blood vessels. However, tumor blood vessels demonstrate a markedly abnormal phenotype that includes several important morphological changes. The degree of angiogenesis is determined by the balance between the angiogenic stimulators and inhibitors released by the tumor and host cells. Recent studies have revealed that TECs also exhibit altered characteristics which depend on the tumor microenvironment. Here, we review recent studies on TEC abnormalities and heterogeneity with respect to tumor progression and consider their therapeutic implications.
• Heterogeneity of tumor endothelial cells and drug delivery

  Kyoko Hida, Nako Maishi, Yu Sakurai, Yasuhiro Hida, Hideyoshi Harashima

  ADVANCED DRUG DELIVERY REVIEWS 99 140 - 147 0169-409X 2016/04 [Refereed][Not invited]
   

  To date anti-angiogenic therapy has been used for cancer therapy widely, yielding promising results. However, it has been elucidated that current anti-angiogenic drug has several issues to be solved, such as side-effects and drug resistance. It has been reported that tumor endothelial cells (TECs) differ from normal counterparts. In addition, it was shown that the TECs are heterogeneous according to the malignancy status of tumor. The development of novel strategy for targeting tumor vasculature is required. Recently, we have developed an active targeting system, which targets TECs specifically. In this review, we will discuss how TECs in tumor vasculature are heterogeneous and offer new perspectives on a drug delivery system, which can target heterogeneous tumor blood vessels from a viewpoint of personalized medicine. (C) 2015 Elsevier B.V. All rights reserved.
• Editorial: Targeting tumor microenvironment heterogeneity

  Kyoko Hida, Genichiro Ishii

  Advanced Drug Delivery Reviews 99 139  1872-8294 2016/04/01 [Refereed][Not invited]
  
• Neuropilin 1 Receptor Is Up-Regulated in Dysplastic Epithelium and Oral Squamous Cell Carcinoma

  Shokoufeh Shahrabi-Farahani, Marina Gallottini, Fabiana Martins, Erik Li, Dayna R. Mudge, Hironao Nakayama, Kyoko Hida, Dipak Panigrahy, Patricia A. D'Amore, Diane R. Bielenberg

  AMERICAN JOURNAL OF PATHOLOGY 186 (4) 1055 - 1064 0002-9440 2016/04 [Refereed][Not invited]
   

  Neuropilins are receptors for disparate ligands, including proangiogenic factors such as vascular endothelial growth factor and inhibitory class 3 semaphorin (SEMA3) family members. Differentiated cells in skin epithelium and cutaneous squamous ceLL carcinoma highly express the neuropilin-1 (NRP1) receptor. We examined the expression of NRP1 in human and mouse oral mucosa. NRP1 was significantly up-regulated in oral epithelial dysplasia and oral squamous cell carcinoma (OSCC). NRP1 receptor localized to the outer suprabasal epithelial layers in normal tongue, an expression pattern similar to the normal skin epidermis. However, dysplastic tongue epithelium and OSCC up regulated NRP1 in basal and proliferating epithelial layers, a profile unseen in cutaneous squamous cell carcinoma. NRP1 up-regulation is observed in a mouse carcinogen-induced OSCC model and in human tongue OSCC biopsies. Human OSCC cell Lines express NRP1 protein in vitro and in mouse tongue xenografts. Sites of capillary infiltration into orthotopic OSCC tumors correlate with high NRP1 expression. HSC3 xenografts, which express the highest NRP1 Levels of the cell lines examined, showed massive intratumoral lymphangiogenesis. SEMA3A inhibited OSCC cell migration, suggesting that the NRP1 receptor was bioactive in OSCC. In conclusion, NRP1 is regulated in the oral epithelium and is selectively up-regulated during epithelial dysplasia. NRP1 may function as a reservoir to sequester proangiogenic ligands within the neoplastic compartment, thereby recruiting neovessels toward tumor cells.
• Comparison of characteristics of mouse immortalized normal endothelial cells, MS1 and primary cultured endothelial cells

  Hida K, Maishi N, Torii C, Yanagiya M, Annan Akuba-Muhyia Dorcas, Morimoto M, Alam Mohammad Towfik

  Hokkaido Journal of Dental Science 北海道歯学会 37 (1) 40 - 48 0914-7063 2016 [Refereed][Not invited]
   

  Tumor blood vessels support the progression of tumors by providing nutrition and oxygen required for growth. By acting as gatekeepers, they allow the metastasis of tumors to secondary locations. An important strategy in cancer therapy has been to target tumor blood vessels consequently inhibiting tumor angiogenesis. To date, antiangiogenic therapy being employed for cancer treatment have yielded profoundly good results. However, it has been shown that current antiangiogenic drugs have several problems, such as adverse side effects and drug resistance. Tumor endothelial cells (TEC), which line the inner layer of blood vessels of the tumor stromal tissue, are the main targets of the antiangiogenic therapies. TEC have been reported to differ significantly from endothelial cells resident in normal blood vessels. These differences provide a window through which TEC can be targeted solely with little or no impact on normal endothelial cells (NEC). Currently, as part of new antiangiogenic drug discovery processes, cell-based screening is being performed using thousands of small chemical compounds. For the success of such screening purposes, there is a need to obtain the right kind of cells and in adequate quantities. Primary–cultured endothelial cells isolated from murine / human blood vessels are the preferred choice. However, maintenance of the primary-cultured endothelial cells is costly and overtime these cells become senescent and perish. As a result, MS1, SV40 immortalized islet-derived endothelial cells, have been used in place of the primary-cultured cells. MS1 is commercially available with comparatively cheaper cell culture requirements. In this study, we compared the characteristics of MS1 and primary-cultured endothelial cells ; NEC and TEC to investigate the possibility of using MS1 cells for chemical screening in search of a new antiangiogenic drug. MS1 cells proliferate faster compared to TEC and upregulated the mRNA expressions of CD133 and Sca-1 genes. However, mRNA expression of most of the other genes, which were upregulated in TEC compared to NEC, were also expressed at lower levels in the MS1 cells. Furthermore, MS1 migrated at a slower rate and did not form tubes on matrigel, as opposed to the function of TEC. In conclusion, MS1 did not completely resemble NEC, nor TEC in function and gene expression. It is suggested that for chemical screening, primary-cultured TEC and the corresponding NEC would be a more ideal choice of cells.
• CXCL12-CXCR7 axis is important for tumor endothelial cell angiogenic property

  Kenji Yamada, Nako Maishi, Kosuke Akiyama, Mohammad Towfik Alam, Noritaka Ohga, Taisuke Kawamoto, Masanobu Shindoh, Norihiko Takahashi, Toshiya Kamiyama, Yasuhiro Hida, Akinobu Taketomi, Kyoko Hida

  INTERNATIONAL JOURNAL OF CANCER 137 (12) 2825 - 2836 0020-7136 2015/12 [Refereed][Not invited]
   

  We reported that tumor endothelial cells (TECs) differ from normal endothelial cells (NECs) in many aspects, such as gene expression profiles. Although CXCR7 is reportedly highly expressed in blood vessels of several tumors, its function in TECs is still unknown. To investigate this role, we isolated TECs from mouse tumor A375SM xenografts, and compared them with NECs from normal mouse dermis. After confirming CXCR7 upregulation in TECs, we analyzed its function using CXCR7 siRNA and CXCR7 inhibitor; CCX771. CXCR7 siRNA and CCX771 inhibited migration, tube formation and resistance to serum starvation in TECs but not in NECs. ERK1/2 phosphorylation was inhibited by CXCR7 knockdown in TECs. These results suggest that CXCR7 promotes angiogenesis in TECs via ERK1/2 phosphorylation. Using ELISA, we also detected CXCL12, a ligand of CXCR7, in conditioned medium from TECs, but not from NECs. CXCL12 neutralizing antibody significantly inhibited TEC random motility. VEGF stimulation upregulated CXCR7 expression in NECs, implying that VEGF mediates CXCR7 expression in endothelial cells. A CXCR7 inhibitor, CCX771 also inhibited tumor growth, lung metastasis and tumor angiogenesis in vivo. Taken together, the CXCL12-CXCR7 autocrine loop affects TEC proangiogenic properties, and could be the basis for an antiangiogenic therapy that specifically targets tumor blood vessels rather than normal vessels. What's new? While the chemokine receptor CXCR7 is expressed at high levels on blood vessels in several tumor types, its function in tumor endothelial cells is unclear. The present study suggests that CXCR7 may have a role in tumor angiogenesis and lung metastasis. CXCR7 and its ligand CXCL12 were found to facilitate tube formation and cell migration in tumor endothelial cells via autocrine signaling. In mice, blockade of CXCR7 with the inhibitory molecule CCX771 resulted in reduced tumor growth and tumor weight, as well as lnhibition of lung metastasis. Inhibition of CXCL12/CXCR7 may represent a promising antiangiogenic strategy.
• Adaptor protein CRK induces epithelial-mesenchymal transition and metastasis of bladder cancer cells through HGF/c-Met feedback loop.

  Ryuji Matsumoto, Masumi Tsuda, Lei Wang, Nako Maishi, Takashige Abe, Taichi Kimura, Mishie Tanino, Hiroshi Nishihara, Kyoko Hida, Yusuke Ohba, Nobuo Shinohara, Katsuya Nonomura, Shinya Tanaka

  Cancer science 106 (6) 709 - 17 2015/06 [Refereed][Not invited]
   

  We have previously reported that an adaptor protein CRK, including CRK-I and CRK-II, plays essential roles in the malignant potential of various aggressive human cancers, suggesting the validity of targeting CRK in molecular targeted therapy of a wide range of cancers. Nevertheless, the role of CRK in human bladder cancer with marked invasion, characterized by distant metastasis and poor prognosis, remains obscure. In the present study, immunohistochemistry indicated a striking enhancement of CRK-I/-II, but not CRK-like, in human bladder cancer tissues compared to normal urothelium. We established CRK-knockdown bladder cancer cells using 5637 and UM-UC-3, which showed a significant decline in cell migration, invasion, and proliferation. It is noteworthy that an elimination of CRK conferred suppressed phosphorylation of c-Met and the downstream scaffold protein Gab1 in a hepatocyte growth factor-dependent and -independent manner. In epithelial-mesenchymal transition-related molecules, E-cadherin was upregulated by CRK elimination, whereas N-cadherin, vimentin, and Zeb1 were downregulated. A similar effect was observed following treatment with c-Met inhibitor SU11274. Depletion of CRK significantly decreased cell proliferation of 5637 and UM-UC-3, consistent with reduced activity of ERK. An orthotopic xenograft model with bioluminescent imaging revealed that CRK knockdown significantly attenuated not only tumor volume but also the number of circulating tumor cells, resulted in a complete abrogation of metastasis. Taken together, this evidence uncovered essential roles of CRK in invasive bladder cancer through the hepatocyte growth factor/c-Met/CRK feedback loop for epithelial-mesenchymal transition induction. Thus, CRK might be a potent molecular target in bladder cancer, particularly for preventing metastasis, leading to the resolution of clinically longstanding critical issues.
• Inhibition of Multidrug Transporter in Tumor Endothelial Cells Enhances Antiangiogenic Effects of Low-Dose Metronomic Paclitaxel

  Kosuke Akiyama, Nako Maishi, Noritaka Ohga, Yasuhiro Hida, Yusuke Ohba, Mohammad Towfik Alam, Taisuke Kawamoto, Hitomi Ohmura, Kenji Yamada, Chisaho Torii, Masanobu Shindoh, Kyoko Hida

  AMERICAN JOURNAL OF PATHOLOGY 185 (2) 572 - 580 0002-9440 2015/02 [Refereed][Not invited]
   

  Tumor angiogenesis plays an important role in tumor progression and metastasis. Tumor endothelial cells (TECs) are a therapeutic target of antiangiogenic chemotherapy that was recently developed and is currently being investigated in the clinic with promising results. Low-dose chemotherapy, which is the Long-term administration of relatively Low doses of chemotherapeutic agents, has been proposed for targeting tumor angiogenesis in various types of cancers. ALthough the efficacy of low-dose chemotherapy has been confirmed in several clinical models, some studies show insufficienttherapeutic effect for malignant cancers. As a possible mechanism of the treatment failure, it has been considered that tumor cells may acquire resistance to this therapy. However, drug resistance by TECs may also be due to another mechanism for resistance of tumor cells to low-dose chemotherapy. We reported elsewhere that TECs were resistant to the anticancer drug paclitaxel, which is a mitotic inhibitor, concomitant with P-glycoprotein up-regulation. Verapamil, a P-glycoprotein inhibitor, abrogated TEC resistance in vitro. Herein, we demonstrated that verapamil coadministration enhanced the effects of Low-dose paclitaxel concomitant with inhibiting tumor angiogenesis in a preclinical in vivo mouse melanoma xenograft model. Furthermore, verapamil coadministration reduced lung metastasis. These results suggest that inhibiting P-gLycoprotein in TECs may be a novel strategy for low-dose chemotherapy targeting TECs.
• Comparative Study of the Sensitivities of Cancer Cells to Doxorubicin, and Relationships between the Effect of the Drug-Efflux Pump P-gp

  Golam Kibria, Hiroto Hatakeyama, Kosuke Akiyama, Kyoko Hida, Hideyoshi Harashima

  BIOLOGICAL & PHARMACEUTICAL BULLETIN 37 (12) 1926 - 1935 0918-6158 2014/12 [Refereed][Not invited]
   

  Multi-drug resistance (MDR) of cancers to chemotherapy including doxorubicin (DOX) is mediated by several factors. To design an effective therapy for the treatment of chemotherapy-resistant cancers, it is essential to explore the elements responsible for mediating MDR. However, exploring these factors in detail in a wide range of tumor types is challenging as several critical analytical steps are involved. Here, we demonstrated the way of exploring the factors mediating MDR in the tumor types without performing the analysis at the molecular level of cells. The sensitivities of 15 different types of cancer cells to DOX were evaluated, and the role of P-glycoprotein (P-gp), one of the major efflux-pumps, was explored. A correlation curve was developed between the intracellular amounts of DOX and the sensitivities of cells, and, based on this correlation, the cells were classified in response to the involvement of P-gp that mediates MDR. P-gp plays an active role in mediating MDR of cancer cells where a correlation between the sensitivities of cells and the accumulated DOX exists. In contrast, in cells that show a resistance to DOX but whose sensitivities are independent of the amount of accumulated drug, it was reasonably presumed that mechanisms other than P-gp are likely to be involved in mediating MDR. Based on the correlation between the availability of a drug and cell sensitivity, it would be reasonable to explore the factors governing cancer MDR, which is essential in designing an effective therapeutic approach for treating chemotherapy-resistant cancers using chemotherapeutic drugs.
• Identification of Tumor Endothelial Cells with High Aldehyde Dehydrogenase Activity and a Highly Angiogenic Phenotype

  Hitomi Ohmura-Kakutani, Kosuke Akiyama, Nako Maishi, Noritaka Ohga, Yasuhiro Hida, Taisuke Kawamoto, Junichiro Iida, Masanobu Shindoh, Kunihiko Tsuchiya, Nobuo Shinohara, Kyoko Hida

  PLOS ONE 9 (12) e113910  1932-6203 2014/12 [Refereed][Not invited]
   

  Tumor blood vessels play an important role in tumor progression and metastasis. It has been reported that tumor endothelial cells (TECs) exhibit highly angiogenic phenotypes compared with those of normal endothelial cells (NECs). TECs show higher proliferative and migratory abilities than those NECs, together with upregulation of vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2). Furthermore, compared with NECs, stem cell markers such as Sca-1, CD90, and multidrug resistance 1 are upregulated in TECs, suggesting that stem-like cells exist in tumor blood vessels. In this study, to reveal the biological role of stem-like TECs, we analyzed expression of the stem cell marker aldehyde dehydrogenase (ALDH) in TECs and characterized ALDH(high) TECs. TECs and NECs were isolated from melanoma-xenografted nude mice and normal dermis, respectively. ALDH mRNA expression and activity were higher in TECs than those in NECs. Next, ALDH(high/low) TECs were isolated by fluorescence-activated cell sorting to compare their characteristics. Compared with ALDH(low) TECs, ALDH(high) TECs formed more tubes on Matrigel-coated plates and sustained the tubular networks longer. Furthermore, VEGFR2 expression was higher in ALDHhigh TECs than that in ALDH(low) TECs. In addition, ALDH was expressed in the tumor blood vessels of in vivo mouse models of melanoma and oral carcinoma, but not in normal blood vessels. These findings indicate that ALDH(high) TECs exhibit an angiogenic phenotype. Stem-like TECs may have an essential role in tumor angiogenesis.
• Suprabasin as a novel tumor endothelial cell marker

  Mohammad T. Alam, Hiroko Nagao-Kitamoto, Noritaka Ohga, Kosuke Akiyama, Nako Maishi, Taisuke Kawamoto, Nobuo Shinohara, Akinobu Taketomi, Masanobu Shindoh, Yasuhiro Hida, Kyoko Hida

  CANCER SCIENCE 105 (12) 1533 - 1540 1347-9032 2014/12 [Refereed][Not invited]
   

  Recent studies have reported that stromal cells contribute to tumor progression. We previously demonstrated that tumor endothelial cells (TEC) characteristics were different from those of normal endothelial cells (NEC). Furthermore, we performed gene profile analysis in TEC and NEC, revealing that suprabasin (SBSN) was upregulated in TEC compared with NEC. However, its role in TEC is still unknown. Here we showed that SBSN expression was higher in isolated human and mouse TEC than in NEC. SBSN knockdown inhibited the migration and tube formation ability of TEC. We also showed that the AKT pathway was a downstream factor of SBSN. These findings suggest that SBSN is involved in the angiogenic potential of TEC and may be a novel TEC marker.
• Ligand density at the surface of a nanoparticle and different uptake mechanism: Two important factors for successful siRNA delivery to liver endothelial cells

  Afsana Akhter, Yasuhiro Hayashi, Yu Sakurai, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  INTERNATIONAL JOURNAL OF PHARMACEUTICS 475 (1-2) 227 - 237 0378-5173 2014/11 [Refereed][Not invited]
   

  The specific delivery of a gene to liver sinusoidal endothelial cells (LSEC) could become a useful strategy for treating various liver diseases associated with such cells. We previously reported that the accumulation of KLGR peptide modified liposomes through liver sinusoidal blood vessels was enhanced after an intravenous administration. Here, we report on an attempt to develop an LSEC targeted nanocarrier system to deliver siRNA for the successful knockdown of LSEC specific gene expression. The system involved the development of a multifunctional envelop-type nano device (MEND) modified with the KLGR peptide for siRNA delivery targeting LSEC. Our developed carrier successfully lowered specific gene expression in LSEC. An in vivo study showed that at a lower density of ligand at the surface of the MEND resulted in the highest knockdown of gene expression in LSEC. This is the first report of the successful delivery of siRNA to LSECs. Further experiments suggest that not only a higher endosomal escape efficiency into the cytosol but also the uptake mechanism as a function of ligand density are two important factors to be considered for targeting LSEC. (C) 2014 Elsevier B.V. All rights reserved.
• Construction of an Aptamer Modified Liposomal System Targeted to Tumor Endothelial Cells

  Mst Naznin Ara, Takashi Matsuda, Mamoru Hyodo, Yu Sakurai, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  BIOLOGICAL & PHARMACEUTICAL BULLETIN 37 (11) 1742 - 1749 0918-6158 2014/11 [Refereed][Not invited]
   

  We describe herein the development of a high affinity and specific DNA aptamer as a new ligand for use in liposomal nanoparticles to target cultured mouse tumor endothelial cells (mTECs). Active targeted nanotechnology based drug delivery systems are currently of great interest, due to their potential for reducing side effects and facilitating the delivery of cytotoxic drugs or genes in a site specific manner. In this study, we report on a promising aptamer candidate AraHH036 that shows selective binding towards mTECs. The aptamer does not bind to normal cells, normal endothelial cells or tumor cells. Therefore, we synthesized an aptamer-polyethylene glycol (PEG) lipid conjugate and prepared aptamer based liposomes (ALPs) by the standard lipid hydration method. First, we quantified the higher capacity of ALPs to internalize into mTECs by incubating ALPs containing 1 mol%, 5 mol% and 10 mol% aptamer of total lipids and compared the results to those for unmodified PEGylated liposomes (PLPs). A confocal laser scanning microscope (CLSM) uptake study indicated that the ALPs were taken up more efficiently than PLPs. The measured K-d value of the ALPs was 142 nM. An intracellular trafficking study confirmed that most of the rhodamine labeled ALPs were taken up and co-localized with the green lysotracker, thus confirming that they were located in lysosomes. Finally, using an aptamer based proteomics approach, the molecular target protein of the aptamer was identified as heat shock protein 70 (HSP70). The results suggest that these ALPs offer promise as a new carrier molecule for delivering anti-angiogenesis drugs to tumor vasculature.
• An aptamer ligand based liposomal nanocarrier system that targets tumor endothelial cells

  Mst Naznin Ara, Takashi Matsuda, Mamoru Hyodo, Yu Sakurai, Hiroto Hatakeyama, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  BIOMATERIALS 35 (25) 7110 - 7120 0142-9612 2014/08 [Refereed][Not invited]
   

  The objective of this study was to construct our recently developed aptamer-modified targeted liposome nano-carrier (Apt-PEG-LPs) system to target primary cultured mouse tumor endothelial cells (mTEC), both in vitro and in vivo. We first synthesized an aptamer-polyethylene glycol 2000-distearoyl phosphoethanolamine (Apt-PEG(2000)-DSPE). The conjugation of the Apt-PEG(2000)-DSPE was confirmed by MALDI-TOF mass spectroscopy. A lipid hydration method was used to prepare Apt-PEG-LPs, in which the outer surface of the PEG-spacer was decorated with the aptamer. Apt-PEG-LPs were significantly taken up by mTECs. Cellular uptake capacity was observed both quantitatively and qualitatively using spectrofluorometry, and confocal laser scanning microscopy (CLSM), respectively. In examining the extent of localization of aptamer-modified liposomes that entered the cells, approximately 39% of the Apt-PEG-LPs were not co-localized with lysotracker, indicating that they had escaped from endosomes. The uptake route involved a receptor mediated pathway, followed by clathrin mediated endocytosis. This Apt-PEG-LP was also applied for in vivo research whether this system could target tumor endothelial cells. Apt-PEG-LP and PEG(5000)-DSPE modified Apt-PEG-LP (Apt/PEG(5000)-LP) were investigated by human renal cell carcinoma (OS-RC-2 cells) inoculating mice using CLSM. Apt-PEG-LP and Apt/PEG(5000)-LP showed higher accumulation on tumor vasculature compared to PEG-LP and the co-localization efficacy of Apt-PEG-LP and Apt/PEG(5000)-LP on TEC were quantified 16% and 25% respectively, which was also better than PEG-LP (3%). The findings suggest that this system is considerable promise for targeting tumor endothelial cells to deliver drugs or genes in vitro and in vivo. (C) 2014 Elsevier Ltd. All rights reserved.
• Identification and expression of troponin T, a new marker on the surface of cultured tumor endothelial cells by aptamer ligand

  Mst. Naznin Ara, Mamoru Hyodo, Noritaka Ohga, Kosuke Akiyama, Kyoko Hida, Yasuhiro Hida, Nobuo Shinohara, Hideyoshi Harashima

  CANCER MEDICINE 3 (4) 825 - 834 2045-7634 2014/08 [Refereed][Not invited]
   

  The identification of a specific biomarker involves the development of new clinical diagnostic tools, and an in-depth understanding of the disease at the molecular level. When new blood vessels form in tumor cells, endothelial cell production is induced, a process that plays a key role in disease progression and metastasis to distinct organs for solid tumor types. The present study reports on the identification of a new biomarker on primary cultured mouse tumor endothelial cells (mTECs) using our recently developed high-affinity DNA aptamer AraHH001 (K-d = 43 nmol/L) assisted proteomics approach. We applied a strategy involving aptamer-facilitated biomarker discovery. Biotin-tagged AraHH001 was incubated with lysates of mTECs and the aptamer-proteins were then conjugated with streptavidin magnetic beads. Finally, the bound proteins were separated by sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. We identified troponin T via matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry, the molecular target of aptamer AraHH001, and its presence was confirmed by measuring mRNA, protein levels, western blot, immunostaining, a gel shift assay of AraHH001 with troponin T. We first report here on the discovery of troponin T on mTECs, a promising and interesting diagnostic tool in the development of antiangiogenic therapy techniques the involves the targeting of the tumor vasculature.
• Identification of novel targets for antiangiogenic therapy by comparing the gene expressions of tumor and normal endothelial cells

  Tsuguteru Otsubo, Yasuhiro Hida, Noritaka Ohga, Hideshi Sato, Toshihiro Kai, Yasushi Matsuki, Hideo Takasu, Kosuke Akiyama, Nako Maishi, Taisuke Kawamoto, Nobuo Shinohara, Katsuya Nonomura, Kyoko Hida

  CANCER SCIENCE 105 (5) 560 - 567 1349-7006 2014/05 [Refereed][Not invited]
   

  Targeting tumor angiogenesis is an established strategy for cancer therapy. Because angiogenesis is not limited to pathological conditions such as cancer, molecular markers that can distinguish between physiological and pathological angiogenesis are required to develop more effective and safer approaches for cancer treatment. To identify such molecules, we determined the gene expression profiles of murine tumor endothelial cells (mTEC) and murine normal endothelial cells using DNA microarray analysis followed by quantitative reverse transcription-polymerase chain reaction analysis. We identified 131 genes that were differentially upregulated in mTEC. Functional analysis using siRNA-mediated gene silencing revealed five novel tumor endothelial cell markers that were involved in the proliferation or migration of mTEC. The expression of DEF6 and TMEM176B was upregulated in tumor vessels of human renal cell carcinoma specimens, suggesting that they are potential targets for antiangiogenic intervention for renal cell carcinoma. Comparative gene expression analysis revealed molecular differences between tumor endothelial cells and normal endothelial cells and identified novel tumor endothelial cell markers that may be exploited to target tumor angiogenesis for cancer treatment.
• Cytoplasmic expression of HuR may be a valuable diagnostic tool for determining the potential for malignant transformation of oral verrucous borderline lesions

  Umma Habiba, Tetsuya Kitamura, Aya Yanagawa-Matsuda, Kyoko Hida, Fumihiro Higashino, Yoichi Ohiro, Yasunori Totsuka, Masanobu Shindoh

  ONCOLOGY REPORTS 31 (4) 1547 - 1554 1021-335X 2014/04 [Refereed][Not invited]
   

  Oral verrucous carcinoma (OVC) is a low grade variant of oral squamous cell carcinoma, and oral verrucous hyperplasia (OVH) is a benign lesion without malignant features. However, pathologists are sometimes presented with borderline lesions and are indecisive as to diagnose them as benign or malignant. Thus, these lesions are tentatively termed oral verrucous lesions (OVLs). HuR is an ARE mRNA-binding protein, normally localized in the nucleus but cytoplasmic exportation is frequently observed in cancer cells. The present study aimed to elucidate whether expression of the HuR protein facilitates the diagnosis of true malignant lesions. Clinicopathological features were evaluated, and immunohistochemical analysis for p53, Ki67 and HuR proteins was performed in 48 cases of OVH, OVC and OVL, and the outcomes were correlated using appropriate statistical analysis. The association of these three proteins in relation to malignant transformation was analyzed after a 3-year follow-up of 25 OVL cases. The basal characteristics (age, gender and location) of all cases had no significant association with the types of lesions. Gingiva (39.4%) was the common site for all lesions. Distribution of the examined proteins had a significant association with the lesions. As compared with the OVLs, the number of immunostained-positive cells was significantly higher in the OVCs and lower in the OVH cases. During follow-up, 24% of the OVLs underwent malignant transformation for which high HuR expression and a diffuse staining pattern in the epithelium were observed. Taken together, the high degree of HuR expression with diffuse staining pattern in the epithelium may be an effective diagnostic tool that determines the potential of OVLs for malignant transformation.
• RNAi-mediated gene knockdown and anti-angiogenic therapy of RCCs using a cyclic RGD-modified liposomal-siRNA system

  Yu Sakurai, Hiroto Hatakeyama, Yusuke Sato, Mamoru Hyodo, Hidetaka Akita, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  JOURNAL OF CONTROLLED RELEASE 173 (10) 110 - 118 0168-3659 2014/01 [Refereed][Not invited]
   

  Angiogenesis is one of crucial processes associated with tumor growth and development, and consequently a prime target for cancer therapy. Although tumor endothelial cells (TECs) play a key role in pathological angiogenesis, investigating phenotypical changes in neovessels when a gene expression in TEC is suppressed is a difficult task. Small interfering RNA (siRNA) represents a potential agent due to its ability to silence a gene of interest. We previously developed a system for in vivo siRNA delivery to cancer cells that involves a liposomal-delivery system, a MEND that contains a unique pH-sensitive cationic lipid, YSK05 (YSK-MEND). In the present study, we report on the development of a system that permits the delivery of siRNA to TECs by combining the YSK-MEND and a ligand that is specific to TECs. Cyclo(Arg-Gly-Asp-D-Phe-Lys) (cRGD) is a well-known ligand to alpha(V)beta(3) integrin, which is selectively expressed at high levels in TECs. We incorporated cRGD into the YSK-MEND (RGD-MEND) to achieve an efficient gene silencing in TECs. Quantitative RT-PCR and the 5' rapid amplification of cDNA ends PCR indicated that the intravenous injection of RGD-MEND at a dose of 4.0 mg/kg induced a significant RNAi-mediated gene reduction in TEC but not in endothelial cells of other organs. Finally, we evaluated the therapeutic potency of the RGD-MEND encapsulating siRNA against vascular endothelial growth factor receptor 2. A substantial delay in tumor growth was observed after three sequential RGD-MEND injections on alternate days. In conclusion, the RGD-MEND represents a new approach for the characterization of TECs and for us in anti-angiogenic therapy. (C) 2013 Elsevier B.V. All rights reserved.
• Heterogeneity of tumor endothelial cells

  Kyoko Hida, Noritaka Ohga, Kosuke Akiyama, Nako Maishi, Yasuhiro Hida

  CANCER SCIENCE 104 (11) 1391 - 1395 1349-7006 2013/11 [Refereed][Not invited]
   

  Tumor blood vessels play important roles in tumor progression and metastasis. Thus, targeting tumor blood vessels is an important strategy for cancer therapy. Tumor endothelial cells (TECs) are the main targets of anti-angiogenic therapy. Although tumor blood vessels generally sprout from pre-existing vessels and have been thought to be genetically normal, they display a markedly abnormal phenotype, including morphological changes. The degree of angiogenesis is determined by the balance between the positive and negative regulating molecules that are released by tumor and host cells in the microenvironment. Reportedly, tumor blood vessels are heterogeneous with TECs differing from normal endothelial cells (in contrast to the conventional view). We recently compared characteristics of different TECs isolated from highly and low metastatic tumors. We found TECs from highly metastatic tumors had more proangiogenic phenotypes than those from low metastatic tumors. Elucidating the variety of TEC phenotypes and identifying TEC molecular signatures should lead to more complete understanding of the mechanisms of tumor progression, discovery of new therapeutic targets, and development of biomarkers. This review considers current studies on TEC heterogeneity and discusses the therapeutic implications of these findings.
• Hypoxia-Induced Reactive Oxygen Species Cause Chromosomal Abnormalities in Endothelial Cells in the Tumor Microenvironment

  Miyako Kondoh, Noritaka Ohga, Kosuke Akiyama, Yasuhiro Hida, Nako Maishi, Alam Mohammad Towfik, Nobuo Inoue, Masanobu Shindoh, Kyoko Hida

  PLOS ONE 8 (11) e80349  1932-6203 2013/11 [Refereed][Not invited]
   

  There is much evidence that hypoxia in the tumor microenvironment enhances tumor progression. In an earlier study, we reported abnormal phenotypes of tumor-associated endothelial cells such as those resistant to chemotherapy and chromosomal instability. Here we investigated the role of hypoxia in the acquisition of chromosomal abnormalities in endothelial cells. Tumor-associated endothelial cells isolated from human tumor xenografts showed chromosomal abnormalities, > 30% of which were aneuploidy. Aneuploidy of the tumor-associated endothelial cells was also shown by simultaneous in-situ hybridization for chromosome 17 and by immunohistochemistry with anti-CD31 antibody for endothelial staining. The aneuploid cells were surrounded by a pimonidazole-positive area, indicating hypoxia. Human microvascular endothelial cells expressed hypoxia-inducible factor 1 and vascular endothelial growth factor A in response to either hypoxia or hypoxia-reoxygenation, and in these conditions, they acquired aneuploidy in 7 days. Induction of aneuploidy was inhibited by either inhibition of vascular endothelial growth factor signaling with vascular endothelial growth factor receptor 2 inhibitor or by inhibition of reactive oxygen species by N-acetyl-L-cysteine. These results indicate that hypoxia induces chromosomal abnormalities in endothelial cells through the induction of reactive oxygen species and excess signaling of vascular endothelial growth factor in the tumor microenvironment.
• Lysyl oxidase secreted by tumour endothelial cells promotes angiogenesis and metastasis

  T. Osawa, N. Ohga, K. Akiyama, Y. Hida, K. Kitayama, T. Kawamoto, K. Yamamoto, N. Maishi, M. Kondoh, Y. Onodera, M. Fujie, N. Shinohara, K. Nonomura, M. Shindoh, K. Hida

  BRITISH JOURNAL OF CANCER 109 (8) 2237 - 2247 0007-0920 2013/10 [Refereed][Not invited]
   

  Background: Molecules that are highly expressed in tumour endothelial cells (TECs) may be candidates for specifically targeting TECs. Using DNA microarray analysis, we found that the lysyl oxidase (LOX) gene was upregulated in TECs compared with its expression in normal endothelial cells (NECs). LOX is an enzyme that enhances invasion and metastasis of tumour cells. However, there are no reports on the function of LOX in isolated TECs. Methods: TECs and NECs were isolated to investigate LOX function in TECs. LOX inhibition of in vivo tumour growth was also assessed using beta-aminopropionitrile (BAPN). Results: LOX expression was higher in TECs than in NECs. LOX knockdown inhibited cell migration and tube formation by TECs, which was associated with decreased phosphorylation of focal adhesion kinase (Tyr 397). Immunostaining showed high LOX expression in human tumour vessels in vivo. Tumour angiogenesis and micrometastasis were inhibited by BAPN in an in vivo tumour model. Conclusion: LOX may be a TEC marker and a possible therapeutic target for novel antiangiogenic therapy.
• Application of POLARIC (TM) fluorophores in an in vivo tumor model

  Nako Maishi, Taisuke Kawamoto, Noritaka Ohga, Koji Yamada, Kosuke Akiyama, Kazuyuki Yamamoto, Takahiro Osawa, Yasuhiro Hida, Kyoko Hida

  ONCOLOGY REPORTS 30 (4) 1695 - 1700 1021-335X 2013/10 [Refereed][Not invited]
   

  Fluorescent and luminescent tools are commonly used to study the dynamics of cancer progression and metastases in real-time. Fluorophores have become essential tools to study biological events. However, few can sustain fluorescence long enough during long-term studies. In the present study, we focused on a series of new amphiphilic fluorophores known as POLARIC (TM), which emit strong fluorescence in lipid bilayers and can be readily modified using the Suzuki-Miyaura cross-coupling reaction. Appropriate chemical modifications of substituent groups can improve target-site specificity, reduce cytotoxicity and prolong emission. Therefore, in contrast to conventional fluorescent probes, these fluorophores show promise for long-term monitoring of biological processes. In the present study, we conducted long-term observations of tumor growth and metastasis using a POLARIC derivative as a novel fluorescent probe. For this purpose, we studied the metastatic melanoma cell line A375-SM, which proliferates at a high rate. We compared the characteristics of the POLARIC probe with the commercially available fluorescent dye PKH26 and fluorescent protein mRFP1. A375-SM cells were labeled with these fluorescent probes and orthotopically implanted into nude mice. The fluorescence emitted by POLARIC was detected more than five weeks after implantation without causing detectable harmful effects on tumor growth. By contrast, fluorescence of cells labeled with PKH26 could not be detected at this same time. Furthermore, POLARIC-, but not PKH26-labeled cells, were also detected in lung metastases. These results indicate that labeling cells with POLARIC fluorophores can significantly extend the time course of in vivo studies on tumor cell growth.
• An Approach to Transgene Expression in Liver Endothelial Cells Using a Liposome-Based Gene Vector Coated with Hyaluronic Acid

  Yuma Yamada, Masahiro Hashida, Yasuhiro Hayashi, Mai Tabata, Mamoru Hyodo, Mst Naznin Ara, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  JOURNAL OF PHARMACEUTICAL SCIENCES 102 (9) 3119 - 3127 0022-3549 2013/09 [Refereed][Not invited]
   

  Dysfunctional sinusoidal liver endothelial cells (LECs) are associated with liver diseases, such as liver fibrosis, cirrhosis, and portal hypertension. Because of this, gene therapy targeted to LECs would be a useful and productive strategy for directly treating these diseases at the level of genes. Here, we report on the development of a transgene vector that specifically targets LECs. The vector is a liposome-based gene vector coated with hyaluronic acid (HA). HA is a natural ligand for LECs and confers desirable properties on particles, rendering them biodegradable, biocompatible, and nonimmunogenic. In this study, we constructed HA-modified carriers, and evaluated cellular uptake and transfection activity using cultured LECs from KSN nude mice (KSN-LECs). Cellular uptake analyses showed that KSN-LECs recognized the HA-modified carriers more effectively than skin endothelial cells. The transfection assay indicated that the efficient gene expression in KSN-LECs, using the HA-modified carriers, required an adequate lipid composition and a functional device to control intracellular trafficking. This finding contributes to our overall knowledge of transgene expression targeted to LECs. (C) 2013 Wiley Periodicals, Inc. and the American Pharmacists Association
• The effect of liposomal size on the targeted delivery of doxorubicin to Integrin alpha v beta 3-expressing tumor endothelial cells

  Golam Kibria, Hiroto Hatakeyama, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  BIOMATERIALS 34 (22) 5617 - 5627 0142-9612 2013/07 [Refereed][Not invited]
   

  Size of the liposomes (LPs) specially governs its biodistribution. In this study, LPs were developed with controlled sizes, where variation in LP size dictates the ligand receptor interaction, cellular internalization and its distribution within the tumor microenvironment. The therapeutic efficacies of doxorubicin (DOX)-loaded RGD modified small size (similar to 100 nm in diameter, dnm) and large size (similar to 300 dnm) PEGylated LPs (RGD-PEG-LPs) were compared to that of Doxil (a clinically used DOX-loaded PEG-LP, similar to 100 dnm) in DOX resistant OSRC-2 (Renal cell carcinoma, RCC) tumor xenografts. Doxil, which accumulated in tumor tissue via the enhanced permeability and retention (EPR) effect, failed to suppress tumor growth. Small size RGD-PEG-LP, that targets the tumor endothelial cells (TECs) and extravasates to tumor cells, failed to provide anti-tumor effect. Large size RGD-PEG-LP preferentially targets the TECs via minimization of the EPR effect, and significantly reduced the tumor growth, which was exerted through its strong anti-angiogenic activity on the tumor vasculature rather than having a direct effect on DOX resistant RCC. The prepared large size RGD-PEG-LP that targets the TECs via interacting with Integrin alpha v beta 3, is a potentially effective and alternate therapeutic strategy for the treatment of DOX resistant tumor cells by utilizing DOX, in cases where Doxil is ineffective. (C) 2013 Elsevier Ltd. All rights reserved.
• Expression of parathyroid hormone-related protein confers malignant potential to mucoepidermoid carcinoma

  Kyosuke Nagamine, Tetsuya Kitamura, Aya Yanagawa-Matsuda, Yoichi Ohiro, Kanchu Tei, Kyoko Hida, Fumihiro Higashino, Yasunori Totsuka, Masanobu Shindoh

  Oncology Reports 29 (6) 2114 - 2118 1021-335X 2013/06 [Refereed][Not invited]
   

  Parathyroid hormone-related protein (PTHrP) is known to induce bone resorption by activating RANKL as well as PTH. PTHrP plays a central role in humoral hypercalcemia, and its expression has been reported to be closely associated with bone metastasis of breast carcinoma. PTHrP expression in oral squamous carcinoma cell lines was investigated, and PTHrP was expressed in oral squamous cell carcinoma cell lines similar to that in a prostate carcinoma cell line. Mucoepidermoid carcinoma is the most common malignant salivary gland tumor composed of different types of cells including a squamous component. Its clinical behavior is highly variable and ranges from slow-growing and indolent to locally aggressive and highly metastatic. We examined the PTHrP expression in mucoepidermoid carcinoma and assessed the significance of its correlation with clinicopathological features. Immunohistochemical detection of PTHrP was carried out in 21 cases of mucoepidermoid carcinoma in the head and neck region. PTHrP was highly detectable in intermediate and epidermoid cells, and abundant expression of PTHrP in intermediate cells had a significant association with cancer malignancy, including lymph node metastasis and/or tumor recurrence. These results suggest that PTHrP expresion can be used as a prognostic factor for mucoepidermoid carcinoma.
• Tumour endothelial cells acquire drug resistance in a tumour microenvironment.

  Kyoko Hida, Kosuke Akiyama, Noritaka Ohga, Nako Maishi, Yasuhiro Hida

  Journal of biochemistry 153 (3) 243 - 9 2013/03 [Refereed][Invited]
   

  Tumour growth is dependent on angiogenesis, and tumour blood vessels are recognized as an important target for cancer therapy. Tumour endothelial cells (TECs) are the main targets of anti-angiogenic therapy. Unlike the traditionally held view, some TECs may be genetically abnormal and might acquire drug resistance. Therefore, we investigated the drug resistance of TECs and the mechanism by which it is acquired. TECs show resistance to paclitaxel through greater mRNA expression of multidrug resistance 1, which encodes P-glycoprotein, as compared with normal endothelial cells. We found that high levels of vascular endothelial growth factor in tumour-conditioned medium may be responsible for upregulated P-glycoprotein expression. This is a novel mechanism for the acquisition of drug resistance by TECs in a tumour microenvironment. This review focuses on the possibility that TECs can acquire drug resistance.
• A liposomal delivery system that targets liver endothelial cells based on a new peptide motif present in the ApoB-100 sequence

  Akhter A, Hayashi Y, Sakurai Y, Ohga N, Hida K, Harashima H

  Int J Pharm Elsevier science bv 456 (1) 195 - 201 0378-5173 2013 [Refereed][Not invited]
   

  Liver dysfunction is associated with a variety of liver diseases, including viral or alcoholic hepatitis, fibrosis, cirrhosis, and portal hypertension. A targeted drug delivery system would be very useful in the treatment of these diseases. We herein describe the development of a system comprised of a new peptide-lipid conjugate for the efficient delivery of molecules to LEC. The RLTRKRGLK sequence (3359-3367), which mediates the association of LDL with arterial CSPG and an LDL receptor, was utilized as a ligand for achieving this goal. The peptide modified PEG-LPs (RLTR-PEG-LPs) were efficiently taken up by primary liver endothelial cells (liver ECs) and other types of cells. In vivo biodistribution and confocal microscopy analysis showed that RLTR-PEG-LPs became widely accumulated in LECs within a short time. Distribution of RLTR-PEG-LPs was greatly reduced with a pretreatment of unlabeled RLTR-PEG-LPs, not cationic LPs, indicating that the sequence is important for LECs. The findings indicate that a reverse sequence of RLTR (KLGR) modified PEG-LPs (KLGR-PEG-LP) did the same pattern compared with RLTR-PEG- LPs, suggesting that the RKR or RXXR sequence might be essential for LECs targeting. Collectively RLTR-PEG-LPs and KLGR-PEG-LPs have the potential for delivering drugs to LECs. (C) 2013 Elsevier B.V. All rights reserved.
• The F-prostaglandin receptor is a novel marker for tumor endothelial cells in renal cell carcinoma

  Kosuke Akiyama, Noritaka Ohga, Nako Maishi, Yasuhiro Hida, Kazuko Kitayama, Taisuke Kawamoto, Takahiro Osawa, Yuko Suzuki, Nobuo Shinohara, Katsuya Nonomura, Masanobu Shindoh, Kyoko Hida

  PATHOLOGY INTERNATIONAL 63 (1) 37 - 44 1320-5463 2013/01 [Refereed][Not invited]
   

  Tumor angiogenesis is necessary for tumor progression and metastasis; therefore, tumor blood vessels are potential therapeutic targets in anticancer therapy. We previously reported that tumor endothelial cells (TECs) exhibit different phenotypes compared with normal endothelial cells (NECs), and microarray analyses of mouse TECs and NECs have shown that several genes are upregulated in TECs compared with NECs. Among these genes, the expression levels of prostaglandin F receptor (PTGFR) mRNA, which encodes the prostaglandin F receptor (FP), were higher in TECs than in NECs. It has been reported that FP and its ligand, prostaglandin F2a, are involved in tumor angiogenesis. However, there have been no reports of the expression of PTGFR in the tumor vessels of renal cell carcinoma (RCC). Thus, we isolated human TECs (hTECs) from RCCs. The expression levels of PTGFR mRNA were also upregulated in hTECs. In addition, immunostaining showed that the PTGFR was expressed in human tumor blood vessels in vivo. These findings suggested that PTGFR is a novel TEC marker and that it may be a novel target for antiangiogenic therapy for RCC.
• Development of a Novel DNA Aptamer Ligand Targeting to Primary Cultured Tumor Endothelial Cells by a Cell-Based SELEX Method

  Mst. Naznin Ara, Mamoru Hyodo, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  PLOS ONE 7 (12) e50174  1932-6203 2012/12 [Refereed][Not invited]
   

  The present study used a spontaneous cell-based SELEX method (Systemic Evolution of Ligands by EXponential Enrichment) to produce DNA aptamers that specifically bind to cell surface proteins or biomarkers produced by primary cultured mouse tumor endothelial cells (mTECs). In solid tumors, new blood vessels are formed through an angiogenesis process, and this plays a critical role in cancer development as well as metastasis. To combat angiogenesis, an appropriate diagnosis and a molecular-level understanding of the different cancer types are now a high priority. The novel DNA aptamer AraHH001, developed in this study, binds specifically to mTECs with high affinity in the nano-molar range, but does not bind to normal skin endothelial cells (skin-ECs). The selected DNA aptamer was also found to bind to cultured human tumor endothelial cells (hTECs), isolated from a clinical patient with a renal carcinoma. The aptamer AraHH001 showed significant anti-angiogenesis activity by inhibiting tube formation by mTECs on matrigel. Interestingly, a confocal laser scanning microscopy examination of in vitro cellular uptake revealed that AraHH001 was assimilated by mTECs, and became co-localized in acidic compartments, as detected by labeling with Lysotracker Red. Therefore, the development of a specific DNA aptamer that binds to mTECs, as reported here for the first time, holds great promise not only as a therapeutic aptamer but also as a targeted molecular probe that appears to play a major role in angiogenesis, and for the development of a targeted new drug delivery system. Citation: Ara MN, Hyodo M, Ohga N, Hida K, Harashima H (2012) Development of a Novel DNA Aptamer Ligand Targeting to Primary Cultured Tumor Endothelial Cells by a Cell-Based SELEX Method. PLoS ONE 7(12): e50174. doi:10.1371/journal.pone.0050174
• Dose swallowing recover in the long term in patients with surgically treated tongue.

  Tei K, Sakakibara N, Yamazaki Y, Ohiro Y, Ono M, Totsuka Y

  Journal of Oral and Maxillofacial Surgery 70 (11) 2680 - 2686 2012/11 [Refereed][Not invited]
  
• Size-controlled, dual-ligand modified liposomes that target the tumor vasculature show promise for use in drug-resistant cancer therapy

  Kazuhiro Takara, Hiroto Hatakeyama, Golam Kibria, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  JOURNAL OF CONTROLLED RELEASE 162 (1) 225 - 232 0168-3659 2012/08 [Refereed][Not invited]
   

  Anti-angiogenic therapy is a potential chemotherapeutic strategy for the treatment of drug resistant cancers. However, a method for delivering such drugs to tumor endothelial cells remains to be a major impediment to the success of anti-angiogenesis therapy. We designed liposomes (LPs) with controlled diameter of around 300 nm, and modified them with a specific ligand and a cell penetrating peptide (CPP) (a dual-ligand LP) for targeting CD13-expressing neovasculature in a renal cell carcinoma (RCC). We modified the LPs with an NGR motif peptide on the top of poly(ethylene glycol) and tetra-arginine (R4) on the surface of the liposome membrane as a specific and CPP ligand, respectively. The large size prevented extravasation of the dual-ligand LP, which allowed it to associate with target vasculature. While a single modification with either the specific or CPP ligand showed no increase in targetability, the dual-ligand enhanced the amount of delivered liposomes after systemic administration to OS-RC-2 xenograft mice. The anti-tumor activity of a dual-ligand LP encapsulating doxorubicin was evaluated and the results were compared with Doxil (R), which is clinically used to target tumor cells. Even though Doxil showed no anti-tumor activity, the dual-ligand LP suppressed tumor growth because the disruption of tumor vessels was efficiently induced. The comparison showed that tumor endothelial cells (TECs) were more sensitive to doxorubicin by 2 orders than RCC tumor cells, and the disruption of tumor vessels was efficiently induced. Collectively, the dual-ligand LP is promising carrier for the treatment of drug resistant RCC via the disruption of TECs. (C) 2012 Elsevier B. V. All rights reserved.
• Prostacyclin receptor in tumor endothelial cells promotes angiogenesis in an autocrine manner

  Takahiro Osawa, Noritaka Ohga, Yasuhiro Hida, Kazuko Kitayama, Kosuke Akiyama, Yuichiro Onodera, Manabu Fujie, Nobuo Shinohara, Masanobu Shindoh, Katsuya Nonomura, Kyoko Hida

  CANCER SCIENCE 103 (6) 1038 - 1044 1347-9032 2012/06 [Refereed][Not invited]
   

  Molecules highly expressed in tumor endothelial cells (TEC) are important for specific targeting of these cells. Previously, using DNA microarray analysis, we found that the prostacyclin receptor (IP receptor) gene was upregulated in TEC compared with normal endothelial cells (NEC). Although prostacyclin is implicated in re-endothelialization and angiogenesis, its role remains largely unknown in TEC. Moreover, the effect of the IP receptor on TEC has not been reported. In the present study we investigated the function of the IP receptor in TEC. The TEC were isolated from two types of human tumor xenografts in nude mice, while NEC were isolated from normal counterparts. Prostacyclin secretion levels in TEC were significantly higher than those in NEC, as shown using ELISA. Real-time RT-PCR showed that the IP receptor was upregulated in TEC compared with NEC. Furthermore, migration and tube formation of TEC were suppressed by the IP receptor antagonist RO1138452. Immunohistostaining showed that the IP receptor was specifically expressed in blood vessels of renal cell carcinoma specimens, but not in glomerular vessels of normal renal tissue. These findings suggest that the IP receptor is a TEC-specific marker and might be a useful therapeutic target. (Cancer Sci 2012; 103: 10381044)
• CXCR7: A novel tumor endothelial marker in renal cell carcinoma

  Nako Maishi, Noritaka Ohga, Yasuhiro Hida, Kosuke Akiyama, Kazuko Kitayama, Takahiro Osawa, Yuichiro Onodera, Nobuo Shinohara, Katsuya Nonomura, Masanobu Shindoh, Kyoko Hida

  PATHOLOGY INTERNATIONAL 62 (5) 309 - 317 1320-5463 2012/05 [Refereed][Not invited]
   

  Tumor angiogenesis is necessary for progression and metastasis of solid tumor. Tumor blood vessels are morphologically different from their normal counterparts. In this study, we isolated tumor endothelial cells (TECs) and revealed their abnormalities. We have compared the gene expression profiles of TECs and normal endothelial cells (NECs) by microarray analysis and found that several genes were upregulated in TECs. Expression of the chemokine receptor CXCR7 mRNA was higher in TECs than in NECs. However, information regarding the expression of CXCR7 in the tumor vessels of renal cell carcinoma is limited. CXCR7 and its ligand CXCL12 have been implicated in tumor cell survival. In this study, the expression of CXCR7 in the tumor vessels of renal cell carcinoma (RCC) was investigated. Real-time PCR revealed higher expression level of CXCR7 in cultured TECs than in cultured NECs. Furthermore, similar to mouse TECs, immunostaining revealed strong expression of CXCR7 in vivo in human tumor vessels. These findings suggest that CXCR7 is a novel TEC marker and a target for antiangiogenic therapy for RCC.
• Tumor Endothelial Cells Acquire Drug Resistance by MDR1 Up-Regulation via VEGF Signaling in Tumor Microenvironment

  Kosuke Akiyama, Noritaka Ohga, Yasuhiro Hida, Taisuke Kawamoto, Yoshihiro Sadamoto, Shuhei Ishikawa, Nako Maishi, Tomoshige Akino, Miyako Kondoh, Aya Matsuda, Nobuo Inoue, Masanobu Shindoh, Kyoko Hida

  AMERICAN JOURNAL OF PATHOLOGY 180 (3) 1283 - 1293 0002-9440 2012/03 [Refereed][Not invited]
   

  Tumor endothelial cells (TECs) are therapeutic targets in anti-angiogenic therapy. Contrary to the traditional assumption, TECs can be genetically abnormal and might also acquire drug resistance. In this study, mouse TECs and normal ECs were isolated to investigate the drug resistance of TECs and the mechanism by which it is acquired. TECs were more resistant to paclitaxel with the up-regulation of multidrug resistance (MDR) 1 mRNA, which encodes the P-glycoprotein, compared with normal ECs. Normal human microvascular ECs were cultured in tumor-conditioned medium (CM) and became more resisting to paclitixel through MDRI mRNA up-regulation and nuclear translocation of Y-box-binding protein 1, which is an MDR1 transcription factor. Vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2) and Akt were activated in human microvascular ECs by tumor CM. We observed that tumor CM contained a significantly high level of VEGF. A VEGFR kinase inhibitor, Ki8751, and a phosphatidylinositol 3-kinase-Akt inhibitor, LY294002, blocked tumor CM-induced MDR1 up-regulation. MDR1 up-regulation, via the VEGF-VEGFR pathway in the tumor microenvironment, is one of the mechanisms of drug resistance acquired by TECs. We observed that VEGF secreted from tumors up-regulated MDR1 through the activation of VEGFR2 and Akt. This process is a novel mechanism of the acquisition of drug resistance by TECs in the tumor microenvironment (Am J Pathol 2012, 180: 1283-1293; DOI: 10.1016/j.ajpath.2011.11.029)
• Heterogeneity of Tumor Endothelial Cells Comparison between Tumor Endothelial Cells Isolated from High- and Low-Metastatic Tumors

  Noritaka Ohga, Shuhei Ishikawa, Nako Maishi, Kosuke Akiyama, Yasuhiro Hida, Taisuke Kawamoto, Yoshihiro Sadamoto, Takahiro Osawa, Kazuyuki Yamamoto, Miyako Kondoh, Hitomi Ohmura, Nobuo Shinohara, Katsuya Nonomura, Masanobu Shindoh, Kyoko Hida

  AMERICAN JOURNAL OF PATHOLOGY 180 (3) 1294 - 1307 0002-9440 2012/03 [Refereed][Not invited]
   

  An important concept in tumor angiogenesis is that tumor endothelial cells (TECs) are genetically normal and homogeneous. However, we previously reported that TECs differ from normal ECs. Whether the characteristics of TECs derived from different tumors differ remains unknown. To elucidate this, in this study, we isolated two types of TECs from high-metastatic (HM) and low-metastatic (LM) tumors and compared their characteristics. HM tumor-derived TECs (HM-TECs) showed higher proliferative activity and invasive activity than LM tumor-derived TECs (LM-TECs). Moreover, the mRNA expression levels of pro-angiogenic genes, such as vascular endothelial growth factor (VEGF) receptors 1 and 2, VEGF, and hypoxia-inducible factor-1 alpha, were higher in HM-TECs than in LM-TECs. The tumor blood vessels themselves and the surrounding area in HM tumors were exposed to hypoxia. Furthermore, HM-TECs showed higher mRNA expression levels of the sternness-related gene stem cell antigen and the mesenchymal marker CD90 compared with LM-TECs. HM-TECs were spheroid, with a smoother surface and higher circularity in the stem cell spheroid assay. HM-TECs differentiated into osteogenic cells, expressing activated alkaline phosphatase in an osteogenic medium at a higher rate than either LM-TECs or normal ECs. Furthermore, HM-TECs contained more aneuploid cells than LM-TECs. These results indicate that TECs from HM tumors have a more pro-angiogenic phenotype than those from LM tumors. (Am J Pathol 2012,180:1294-1307; DOI: 10.1016/j.ajpath.2011.11.035)
• Biglycan is a specific marker and an autocrine angiogenic factor of tumour endothelial cells

  K. Yamamoto, N. Ohga, Y. Hida, N. Maishi, T. Kawamoto, K. Kitayama, K. Akiyama, T. Osawa, M. Kondoh, K. Matsuda, Y. Onodera, M. Fujie, K. Kaga, S. Hirano, N. Shinohara, M. Shindoh, K. Hida

  BRITISH JOURNAL OF CANCER 106 (6) 1214 - 1223 0007-0920 2012/03 [Refereed][Not invited]
   

  BACKGROUND: We isolated tumour endothelial cells (TECs), demonstrated their abnormalities, compared gene expression profiles of TECs and normal endothelial cells (NECs) by microarray analysis and identified several genes upregulated in TECs. We focused on the gene encoding biglycan, a small leucine-rich repeat proteoglycan. No report is available on biglycan expression or function in TECs. METHODS: The NEC and TEC were isolated. We investigated the biglycan expression and function in TECs. Western blotting analysis of biglycan was performed on sera from cancer patients. RESULTS: Biglycan expression levels were higher in TECs than in NECs. Biglycan knockdown inhibited cell migration and caused morphological changes in TECs. Furthermore, immunostaining revealed strong biglycan expression in vivo in human tumour vessels, as in mouse TECs. Biglycan was detected in the sera of cancer patients but was hardly detected in those of healthy volunteers. CONCLUSION: These findings suggested that biglycan is a novel TEC marker and a target for anti-angiogenic therapy. British Journal of Cancer (2012) 106, 1214-1223. doi: 10.1038/bjc.2012.59 www.bjcancer.com Published online 28 February 2012 (C) 2012 Cancer Research UK
• Tumor-Derived Microvesicles Induce Proangiogenic Phenotype in Endothelial Cells via Endocytosis

  Taisuke Kawamoto, Noritaka Ohga, Kosuke Akiyama, Naoya Hirata, Shuji Kitahara, Nako Maishi, Takahiro Osawa, Kazuyuki Yamamoto, Miyako Kondoh, Masanobu Shindoh, Yasuhiro Hida, Kyoko Hida

  PLOS ONE 7 (3) e34045  1932-6203 2012/03 [Refereed][Not invited]
   

  Background: Increasing evidence indicates that tumor endothelial cells (TEC) differ from normal endothelial cells (NEC). Our previous reports also showed that TEC were different from NEC. For example, TEC have chromosomal abnormality and proangiogenic properties such as high motility and proliferative activity. However, the mechanism by which TEC acquire a specific character remains unclear. To investigate this mechanism, we focused on tumor-derived microvesicles (TMV). Recent studies have shown that TMV contain numerous types of bioactive molecules and affect normal stromal cells in the tumor microenvironment. However, most of the functional mechanisms of TMV remain unclear. Methodology/Principal Findings: Here we showed that TMV isolated from tumor cells were taken up by NEC through endocytosis. In addition, we found that TMV promoted random motility and tube formation through the activation of the phosphoinositide 3-kinase/Akt pathway in NEC. Moreover, the effects induced by TMV were inhibited by the endocytosis inhibitor dynasore. Our results indicate that TMV could confer proangiogenic properties to NEC partly via endocytosis. Conclusion: We for the first time showed that endocytosis of TMV contributes to tumor angiogenesis. These findings offer new insights into cancer therapies and the crosstalk between tumor and endothelial cells mediated by TMV in the tumor microenvironment.
• Cyclooxygenase-2 inhibition causes antiangiogenic effects on tumor endothelial and vascular progenitor cells

  Chikara Muraki, Noritaka Ohga, Yasuhiro Hida, Hiroshi Nishihara, Yasutaka Kato, Kunihiko Tsuchiya, Kohei Matsuda, Yasunori Totsuka, Masanobu Shindoh, Kyoko Hida

  INTERNATIONAL JOURNAL OF CANCER 130 (1) 59 - 70 0020-7136 2012/01 [Refereed][Not invited]
   

  Tumor angiogenesis is necessary for solid tumor progression and metastasis. Cyclooxygenase (COX)-2 is known to play an important role in cancer growth and invasion, and it activates the signaling pathways controlling cell proliferation, migration, apoptosis, and angiogenesis. COX-2 is reported to be expressed in many cancer cells. Several studies have reported successful treatment of cancer cells with COX-2 inhibitors (COX-2is). However, the effect of COX-2 inhibition on the tumor endothelium remains to be elucidated. Our study shows that COX-2 is expressed in the vasculature of surgically resected human tumors. To investigate the effects of COX-2 inhibition on the tumor endothelium in vitro, we isolated tumor endothelial cells (TECs) from human melanoma and oral carcinoma xenografts in mice, in which we confirmed that tumor growth was suppressed by inhibiting angiogenesis with the COX-2is NS398. COX-2 mRNA was upregulated in TECs compared to normal endothelial cells (NECs). Cell migration and proliferation were suppressed by NS398 in TECs but not in NECs. The effects of NS398 in vivo were consistent with the in vitro results. The number of CD133(+)/vascular endothelial growth factor receptor-2(+) cells in circulation was significantly suppressed by COX-2 inhibition. In addition, the number of progenitor marker-positive cells decreased in the tumor blood vessels after COX-2i treatment, which suggests that the homing of progenitor cells into the tumor was also blocked. We conclude that NS398 specifically targets both TECs and vascular progenitor cells without affecting NECs.
• Altered angiogenesis in the tumor microenvironment

  Kyoko Hida, Taisuke Kawamoto, Noritaka Ohga, Kosuke Akiyama, Yasuhiro Hida, Masanobu Shindoh

  PATHOLOGY INTERNATIONAL 61 (11) 630 - 637 1320-5463 2011/11 [Refereed][Not invited]
   

  Tumor blood vessels play an important role in tumor progression and metastasis. Thus, targeting the tumor blood vessels is an important strategy in cancer therapy. Tumor blood vessels generally arise from pre-existing vessels and have been thought to be genetically normal. However, they have been shown to differ from their normal counterparts, e. g. with regard to the morphological changes. We isolated tumor endothelial cells (TEC) from mouse tumor xenografts and showed that they were abnormal. TEC up-regulate many genes, proliferate more rapidly and migrate more than normal endothelial cells (NEC). Furthermore, the TEC in our study were cytogenetically abnormal. We concluded that TEC can acquire cytogenetic abnormalities while in the tumor microenvironment. In order to develop ideal antiangiogenic therapies, understanding the crosstalk between blood vessels and the tumor microenvironment is important. This review considers the current studies on TEC abnormalities and discusses the possible mechanism by which the tumor microenvironment produces abnormal TEC.
• Dual-ligand modification of PEGylated liposomes shows better cell selectivity and efficient gene delivery

  Golam Kibria, Hiroto Hatakeyama, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  JOURNAL OF CONTROLLED RELEASE 153 (2) 141 - 148 0168-3659 2011/07 [Refereed][Not invited]
   

  The objective of this study was to develop an efficient dual-ligand based PEGylated liposomal delivery system that had target specificity as well as properties that would enhance cellular uptake. PEGylated liposomes (PEG-LP) were prepared by the lipid film hydration method by adding distearoyl phosphoethanolamine-polyethylene-glycol-2000 conjugate (DSPE-PEG2000) to a lipid mixture. The cyclic RGD (Arg-Gly-Asp) peptide, a specific ligand with affinity for Integrin alpha(v)beta(3) was coupled to the distal end of the PEG on the PEG-LP (RGD-PEG-LP). Stearylated octaarginine (STR-R8) was incorporated on the surface of the RGD-PEG-LP as dual-ligand (R8/RGD-PEG-LP) that functions as a cell penetrating peptide (CPP). RGD-PEG-LP and R8/RGD-PEG-LP were preferentially taken up by caveolae-mediated and clathrin-mediated endocytosis pathways, respectively. Compared to PEG-LP, R8/RGD-PEG-LP showed an enhanced cellular uptake as well as a higher transfection efficiency in Integrin alpha(v)beta(3) expressing cells. However, the amount of cellular uptake or gene expression by the single ligand versions was negligible, even in Integrin alpha(v)beta(3) expressing cells. No remarkable difference in cellular uptake or gene expression was observed for cells in which the expression of targeted receptors was absent. It can be concluded that dual-ligand modified PEG-LP possesses a strong capability for the efficient internalization of PEG-LP and consequently would be an effective tool for the targeted delivery of macromolecules or chemotherapeutics through accelerated cellular uptake. (C) 2011 Elsevier B.V. All rights reserved.
• HuR keeps an angiogenic switch on by stabilising mRNA of VEGF and COX-2 in tumour endothelium

  T. Kurosu, N. Ohga, Y. Hida, N. Maishi, K. Akiyama, W. Kakuguchi, T. Kuroshima, M. Kondo, T. Akino, Y. Totsuka, M. Shindoh, F. Higashino, K. Hida

  BRITISH JOURNAL OF CANCER 104 (5) 819 - 829 0007-0920 2011/03 [Refereed][Not invited]
   

  BACKGROUND: Tumour stromal cells differ from its normal counterpart. We have shown that tumour endothelial cells (TECs) isolated from tumour tissues are also abnormal. Furthermore, we found that mRNAs of vascular endothelial growth factor-A (VEGF-A) and cyclooxygenase-2 (COX-2) were upregulated in TECs. Vascular endothelial growth factor-A and COX-2 are angiogenic factors and their mRNAs contain an AU-rich element (ARE). AU-rich element-containing mRNAs are reportedly stabilised by Hu antigen R (HuR), which is exported to the cytoplasm. METHODS: Normal endothelial cell (NEC) and two types of TECs were isolated. We evaluated the correlation of HuR and accumulation of VEGF-A and COX-2 mRNAs in TECs and effects of HuR on biological phenotypes of TECs. RESULTS: The HuR protein was accumulated in the cytoplasm of TECs, but not in NECs. Vascular endothelial growth factor-A and COX-2 mRNA levels decreased due to HuR knockdown and it was shown that these ARE-mRNA were bound to HuR in TECs. Furthermore, HuR knockdown inhibited cell survival, random motility, tube formation, and Akt phosphorylation in TECs. CONCLUSION: Hu antigen R is associated with the upregulation of VEGF-A and COX-2 mRNA in TECs, and has an important role in keeping an angiogenic switch on, through activating angiogenic phenotype in tumour endothelium. British Journal of Cancer (2011) 104, 819-829. doi:10.1038/bjc.2011.20 www.bjcancer.com Published online 1 February 2011 (C) 2011 Cancer Research UK
• 腫瘍血管内皮細胞におけるHuRを介したCOX2およびVEGF-AのmRNAの発現亢進

  黒須 拓郎, 大賀 則孝, 東野 史裕, 格口 渉, 黒嶋 雄志, 秋山 廣輔, 近藤 美弥子, 間石 奈湖, 川本 泰輔, 戸塚 靖則, 進藤 正信, 樋田 京子

  日本口腔科学会雑誌 (NPO)日本口腔科学会 60 (1) 77 - 77 0029-0297 2011/01
• Phenotype of tumor lymphatic vessels is a prognostic factor in human tongue squamous cell carcinoma

  Yukie Nitta, Kyoko Hida, Tetsuya Kitamura, Fumihiro Higashino, Noritaka Ohga, Kazuaki Fukushima, Masanobu Shindoh

  ONCOLOGY LETTERS 2 (1) 79 - 83 1792-1074 2011/01 [Refereed][Not invited]
   

  Tumor metastasis to lymph nodes occurs through the lymphatic vessels located in the tumor circumference. However, few studies have focused on the phenotypes of lymphatic vessels around these tumors. We investigated the characteristics of the lymph vessels of tongue squamous cell carcinoma (SCC) and compared them to clinicopathological characteristics. A total of 43 patients diagnosed as having tongue SCC consulted Hokkaido University Hospital were examined. The lymphatic vessels were identified by antibody D2-40 and the number and diameter of tumor lymphatic vessels were measured. The proliferative activity of lymphatic endothelial cells was also examined by immunostaining using antibody MIB-1. We then measured the DNA density of lymphatic endothelial cells in normal and tumor tissues. The number of tumor lymphatic vessels significantly increased in highly metastatic cases of tongue SCC, particularly in cases with a large number of micro lymphatic vessels. A significant correlation was found between the metastatic and proliferative activity of tumor lymphatic endothelial cells. Moreover, the DNA density of tumor lymphatic endothelial cells increased compared to normal tissues. These results suggest that the phenotypes of tumor lymphatic endothelial cells are an indicator of lymph node metastasis of tongue SCC.
• Design of a dual-ligand system using a specific ligand and cell penetrating peptide, resulting in a synergistic effect on selectivity and cellular uptake

  Kazuhiro Takara, Hiroto Hatakeyama, Noritaka Ohga, Kyoko Hida, Hideyoshi Harashima

  INTERNATIONAL JOURNAL OF PHARMACEUTICS 396 (1-2) 143 - 148 0378-5173 2010/08 [Refereed][Not invited]
   

  In this study, a dual-ligand liposomal system comprised of a specific ligand and a cell penetrating peptide (CPP) is described to enhance selectivity and cellular uptake. Dual-ligand PEGylated liposomes were prepared by modifying the end of the PEG with an NGR motif peptide, followed by a surface coating of the liposomes with stearylated oligoarginine (STR-RX) The NGR motif recognizes CD13. a marker protein located on tumor endothelial cells A suitable number of RX units was determined to be R4, since it can be masked by the PEG aqueous layer Although no enhanced cellular uptake was observed when a single modification of PEGylated liposomes with either NGR- or STR-R4 was used, the dual-modification with NGR and STR-R4 stimulated uptake of PEGylated liposomes by CD13 positive cells, and this uptake was superior to that obtained by PEG-unmodified liposomes modified with STR-R4 The dual-ligand system shows a synergistic effect on cellular uptake Collectively, the dual-ligand system promises to be useful in the development efficient and specific drug delivery systems. (C) 2010 Elsevier B V All rights reserved.
• Adrenomedullin antagonist suppresses tumor formation in renal cell carcinoma through inhibitory effects on tumor endothelial cells and endothelial progenitor mobilization

  Kunihiko Tsuchiya, Kyoko Hida, Yasuhiro Hida, Chikara Muraki, Noritaka Ohga, Tomoshige Akino, Takeshi Kondo, Tetsuya Miseki, Koji Nakagawa, Masanobu Shindoh, Toru Harabayashi, Nobuo Shinohara, Katsuya Nonomura, Masanobu Kobayashi

  INTERNATIONAL JOURNAL OF ONCOLOGY 36 (6) 1379 - 1386 1019-6439 2010/06 [Refereed][Not invited]
   

  Adrenomedullin (AM) is a multifunctional 52-amino acid peptide. AM has several effects and acts as a growth factor in several types of cancer cells. Our previous study revealed that an AM antagonist (AMA) suppressed the growth of pancreatic tumors in mice, although its mechanism was not elucidated. In this study, we constructed an AMA expression vector and used it to treat renal cell carcinoma (RCC) in mice. This AMA expression vector significantly reduced tumor growth in mice. In addition, microvessel density was decreased in AMA-treated tumors. To analyze the effect of AMA on tumor angiogenesis in this model, tumor endothelial cells (TECs) were isolated from RCC xenografts. TEC proliferation was stimulated by AM and it was inhibited by AMA significantly. AM induced migration of TECs and it was also blocked by AMA. However, normal ECs (NECs) were not affected by either AM or AMA. These results demonstrate that AMA has inhibitory effects on TECs specifically, not on NEC, thereby inhibiting tumor angiogenesis. Furthermore, we showed that vascular endothelial growth factor-induced mobilization of endothelial progenitor cell (EPC) into circulation was inhibited by AMA. These results suggest that AMA can be considered a good anti-angiogenic reagent that selectively targets TECs and EPC in renal cancer.
• BMP-9 induces proliferation of multiple types of endothelial cells in vitro and in vivo

  Yuka Suzuki, Noritaka Ohga, Yasuyuki Morishita, Kyoko Hida, Kohei Miyazono, Tetsuro Watabe

  JOURNAL OF CELL SCIENCE 123 (10) 1684 - 1692 0021-9533 2010/05 [Refereed][Not invited]
   

  Members of the bone morphogenetic protein (BMP) family have been implicated in the development and maintenance of vascular systems. Whereas members of the BMP-2/4 and osteogenic protein-1 groups signal via activin receptor-like kinase (ALK)-2, ALK-3 and ALK-6, BMP-9 and BMP-10 have been reported to bind to ALK-1 in endothelial cells. However, the roles of BMP-9-ALK-1 signaling in the regulation of endothelial cells have not yet been fully elucidated. Here, using various systems, we examined the effects of BMP-9 on the proliferation of endothelial cells. Vascular-tube formation from ex vivo allantoic explants of mouse embryos was promoted by BMP-9. BMP-9, as well as BMP-4 and BMP-6, also induced the proliferation of in-vitro-cultured mouse embryonic-stem-cell-derived endothelial cells (MESECs) by inducing the expression of vascular endothelial growth factor receptor 2 and Tie2, a receptor for angiopoietin-1. A decrease in ALK-1 expression or expression of constitutively active ALK-1 in MESECs abrogated and mimicked the effects of BMP-9 on the proliferation of MESECs, respectively, suggesting that BMP-9 promotes the proliferation of these cells via ALK-1. Furthermore, in vivo angiogenesis was promoted by BMP-9 in a Matrigel plug assay and in a BxPC3 xenograft model of human pancreatic cancer. Consistent with these in vivo findings, BMP-9 enhanced the proliferation of in-vitro-cultured normal endothelial cells from dermal tissues of adult mice and of tumor-associated endothelial cells isolated from tumor xenografts in host mice. These findings suggest that BMP-9 signaling activates the endothelium tested in the present study via ALK-1.
• Isolation and culture of microvascular endothelial cells from murine inguinal and epididymal adipose tissues

  Kazuaki Kajimoto, Md. Nazir Hossen, Kyoko Hida, Noritaka Ohga, Hidetaka Akita, Mamoru Hyodo, Yasuhiro Hida, Hideyoshi Harashima

  JOURNAL OF IMMUNOLOGICAL METHODS 357 (1-2) 43 - 50 0022-1759 2010/05 [Refereed][Not invited]
   

  Adipose tissue has long been considered to be a simple tissue that contains adipocytes. Because of this, the isolation and characterization of microvascular endothelical cells, which are also present in adipose tissue, have been neglected, even though they are components of a capillary network that surrounds each individual adipocyte. Here we report on a protocol for producing highly purified murine microvascular endothelial cells (MECs) from diverse sites of murine adipose tissues including inguinal and epididymal adipose tissues. The method is based on a combination of negative and positive immunomagnetic selection. The protocol involves the preparation of a single cell suspension (digestion, filtration and density gradient centrifugation), immunomagnetic enrichment of the CD45(-) cell population and the purification of the MECs by a combination of common specific markers CD31, CD 102 and isolectin B4. The isolated MECs can be successively cultured for 10 to 12 passages without any detectable changes in morphology and phenotype. Therefore, the method described herein represents a protocol for the isolation and long-term maintenance of highly pure mouse MECs in high yields from adipose tissues. (C) 2010 Elsevier B.V. All rights reserved.
• Isolated tumor endothelial cells maintain specific character during long-term culture

  Kohei Matsuda, Noritaka Ohga, Yasuhiro Hida, Chikara Muraki, Kunihiko Tsuchiya, Takuro Kurosu, Tomoshige Akino, Shou-Ching Shih, Yasunori Totsuka, Michael Klagsbrun, Masanobu Shindoh, Kyoko Hida

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 394 (4) 947 - 954 0006-291X 2010/04 [Refereed][Not invited]
   

  Tumor angiogenesis is necessary for solid tumor progression and metastasis. Increasing evidence indicates that tumor endothelial cells (TECs) are more relevant to the study of tumor angiogenesis than normal endothelial cells (NECs) because their morphologies and gene expression are different from NECs. However, it is challenging to isolate and culture large numbers of pure ECs from tumor tissue since the percentage of ECs is only about 1-2% and tumor cells and fibroblasts easily overgrow them. In addition, there has been concern that isolated TECs may lose their special phenotype once they are dissociated from tumor cells. In this study, we have successfully purified murine TECs from four different human tumor xenografts and NECs from murine dermal tissue. Isolated ECs expressed endothelial markers, such as CD31, VE-cadherin (CD144), and endoglin (CD105), for more than 3 months after isolation. TECs maintained tumor endothelial-specific markers, such as tumor endothelial marker 8 (TEM8) and aminopeptidase N (APN), as in tumor blood vessels in vivo. In addition, TECs were more proliferative and motile than NECs. TECs showed a higher response to VEGF and higher expression of VEGF receptors-1 and -2 than NECs did. Stem cell antigen-1 was up-regulated in all four TECs, suggesting that they have a kind of sternness. Cultured TECs maintain distinct biological differences from NECs as in vivo. In conclusion, it was suggested that TECs are relevant material for tumor angiogenesis research. (C) 2010 Elsevier Inc. All rights reserved.
• Significance of anti-angiogenic therapy in head and neck cancer-Heterogeneity of tumor endothelium

  Kyoko Hida, Noritaka Ohga, Yasuhiro Hida, Masanobu Shindoh

  Japanese Dental Science Review 46 (1) 26 - 32 1882-7616 2010/02 [Refereed][Not invited]
   

  Tumor angiogenesis is necessary for solid tumor progression and metastasis. Thus, targeting tumor blood vessels is an important strategy for cancer therapy. Especially, it would give large benefit to head and neck cancer patients if ideal anti-angiogenic drug is developed. Tumor blood vessels have been shown to differ from their normal counterparts, for example, by changes in morphology. An important concept in tumor angiogenesis is that tumor endothelial cells are assumed to be genetically normal, even though these endothelial cells are structurally and functionally abnormal. To date, many anti-angiogenic drugs have been developed, but it has been also reported to cause toxic side effects. To develop ideal anti-angiogenic therapies, understanding tumor endothelial cell abnormalities is important. We have isolated tumor endothelial cells from mouse tumor xenografts and have shown that tumor endothelial cells are abnormal. Tumor endothelial cells upregulate many genes, such as epidermal growth factor receptor. Tumor endothelial cells are also more sensitive to EGF. Unexpectedly, tumor endothelial cells were cytogenetically abnormal. In marked contrast, freshly isolated normal endothelial cells were diploid. We conclude that tumor endothelial cells can acquire cytogenetic abnormalities while in the tumor microenvironment. Here, we provide an overview of the current studies on tumor endothelial cell abnormalities. © 2009 Japanese Association for Dental Science.
• Cytogenetic Abnormalities of Tumor-Associated Endothelial Cells in Human Malignant Tumors

  Tomoshige Akino, Kyoko Hida, Yasuhiro Hida, Kunihiko Tsuchiya, Deborah Freedman, Chikara Muraki, Noritaka Ohga, Kouhei Matsuda, Kousuke Akiyama, Toru Harabayashi, Nobuo Shinohara, Katsuya Nonomura, Michael Klagsbrun, Masanobu Shindoh

  AMERICAN JOURNAL OF PATHOLOGY 175 (6) 2657 - 2667 0002-9440 2009/12 [Refereed][Not invited]
   

  Tumor blood vessels are thought to contain genetically normal and stable endothelial cells (ECs), unlike tumor cells, which typically display genetic instability. Yet, chromosomal aberration in human tumor-associated ECs (hTECs) in carcinoma has not yet been investigated. Here we isolated TECs from 20 human renal cell carcinomas and analyzed their cytogenetic abnormalities. The degree of aneuploidy was analyzed by fluorescence in situ hybridization using chromosome 7 and chromosome 8 DNA probes in isolated hTECs. In human renal cell carcinomas, 22-58% (median, 33%) of uncultured hTECs were aneuploid, whereas normal ECs were diploid. The mechanisms governing TEC aneuploidy were then studied using mouse TECs (mTECs) isolated from xenografts of human epithelial tumors. To investigate the contribution of progenitor cells to aneuploidy in mTECs, CD133(+) and CD133(-) mTECs were compared for aneuploidy. CD133+ mTECs showed aneuploidy more frequently than CD133- mTECs. This is the first report showing cytogenetic abnormality of hTECs in carcinoma, contrary to traditional belief. Cytogenetic alterations in tumor vessels of carcinoma therefore can occur and may play a significant role in modifying tumor-stromal interactions. (Am J Pathol 2009, 175:2657-2667; DOI: 10.2353/ajpath.2009.090202)
• Inhibitory effects of epigallocatechin-3 gallate, a polyphenol in green tea, on tumor-associated endothelial cells and endothelial progenitor cells.

  Noritaka Ohga, Kyoko Hida, Yasuhiro Hida, Chikara Muraki, Kunihiko Tsuchiya, Kohei Matsuda, Yoichi Ohiro, Yasunori Totsuka, Masanobu Shindoh

  Cancer science 100 (10) 1963 - 70 1347-9032 2009/10 [Refereed][Not invited]
   

  The polyphenol epigallocatechin-3 gallate (EGCG) in green tea suppresses tumor growth by direct action on tumor cells and by inhibition of angiogenesis, but it is not known whether it specifically inhibits tumor angiogenesis. We examined the anti-angiogenic effect of EGCG on tumor-associated endothelial cells (TEC), endothelial progenitor cells (EPC), and normal endothelial cells (NEC). EGCG suppressed the migration of TEC and EPC but not NEC. EGCG also inhibited the phosphorylation of Akt in TEC but not in NEC. Furthermore, vascular endothelial growth factor-induced mobilization of EPC into circulation was inhibited by EGCG. MMP-9 in the bone marrow plasma plays key roles in EPC mobilization into circulation. We observed that expression of MMP-9 mRNA was downregulated by EGCG in mouse bone marrow stromal cells. In an in vivo model, EGCG suppressed growth of melanoma and reduced microvessel density. Our study showed that EGCG has selective anti-angiogenic effects on TEC and EPC. It is suggested that EGCG could be a promising angiogenesis inhibitor for cancer therapy.
• 腫瘍血管内皮細胞におけるHuRを介したCOX2およびVEGF-AのmRNAの発現亢進

  黒須 拓郎, 大賀 則孝, 格口 渉, 黒嶋 雄志, 戸塚 靖則, 進藤 正信, 樋田 京子

  日本口腔外科学会雑誌 (公社)日本口腔外科学会 55 (Suppl.) 93 - 93 0021-5163 2009/09
• Pim-1 activation of cell motility induces the malignant phenotype of tongue carcinoma

  Souichi Tanaka, Tetsuya Kitamura, Fumihiro Higashino, Kyoko Hida, Yoichi Ohiro, Mitsunobu Ono, Masanobu Kobayashi, Yasunori Totsuka, Masanobu Shindoh

  MOLECULAR MEDICINE REPORTS 2 (2) 313 - 318 1791-2997 2009/03 [Refereed][Not invited]
   

  Pim-1 is a serine/threonine kinase as well as a protooncogene that induces T-cell lymphoma. Pim-1 induces cell cycle progression in cooperation with c-Myc and acts as an inhibitor of apoptotic cell death, actions that are involved in blood cell oncogenesis. However, little is known regarding the role of Pim-1 in oral carcinogenesis. We investigated Pim-1 expression in tongue squamous cell carcinoma (SCC) and examined its clinicopathological features. Western blotting was performed in 6 oral SCC cell lines, with Pim-1 being detected in all 6 of the lines. Immunohistochemical detection of Pim-1 was carried out in 39 cases of tongue SCC and analyzed in terms of its associated clinicopathological features. Pim-1 was expressed in 17/39 cases of tongue carcinoma, and was significantly correlated with lymph node metastasis. The role of Pim-1 in cell motility was further examined in HSC3 cells using the GTP-binding assay for Rho family protein, the motility assay and siRNA treatment. Rac 1 activation was observed, and cell motility was reduced when Pim-1 was knocked down by siRNA. These results indicate. that Pim-1 is involved in the carcinogenesis of oral SCC and is correlated to metastasis, which is in part associated with the enhancement of cell motility.
• Crosstalk between Blood Vessels and Tumor Microenvironment

  Kyoko Hida, Noritaka Ohga, Takuro Kurosu, Yasunori Totsuka, Masanobu Shindoh

  Oral Science International 7 (1) 1 - 10 1348-8643 2009 [Refereed][Not invited]
   

  Tumor blood vessels play an important role in tumor progression and metastasis. Thus, targeting tumor blood vessels is an important strategy for cancer therapy, especially for head and neck cancer patients. Tumor blood vessels generally sprout from pre-existing vessels and have been thought to be genetically normal. However, tumor blood vessels have been shown to differ from their normal counterparts, for example, by changes in morphology. The authors isolated tumor endothelial cells (TECs) from mouse tumor xenografts and have shown that the TECs are abnormal. TECs up-regulate many genes and proliferate more rapidly and migrate more than normal endothelial cells (NECs). Furthermore, TECs were found to be cytogenetically abnormal. We conclude that TECs can acquire cytogenetic abnormalities while in a tumor microenvironment. To develop ideal antiangiogenic therapies, understanding the crosstalk between blood vessels and the tumor microenvironment is important. Here, we provide an overview of the current studies on TEC abnormalities and a discussion about possible mechanisms for how tumor the microenvironment makes TECs abnormal. © 2009, Japanese Stomatological Society. All rights reserved.
• Aberrant Control of Transcription and Transportation of Oncogenic mRNA in Carcinogenesis

  Fumihiro Higashino, Kyoko Hida, Tetsuya Kitamura, Noritaka Ohga, Masanobu Shindoh

  Oral Science International 7 (1) 11 - 18 1348-8643 2009 [Refereed][Not invited]
   

  Multiple gene disorders have been shown to be involved in carcinogenesis. Mutation, translocation and amplification have been identified in so-called oncogenes, and inactivation of antioncogenes by mutation and deletion has been shown. EIAF is an ets-oncogene family transcription factor, and has been shown to upregulate multiple matrix metalloproteinase (MMP) genes that contribute to the malignant phenotype of cancer cells by inducing invasive and metastatic activities. EWS/ETS fusions are frequently observed in Ewing's sarcoma, and we have revealed that EWS/ETS chimeric protein activates telomerase activity by upregulating human telomerase reverse transcriptase (hTERT), but the transcriptional activation of hTERT by EWS/ETS was indirect, and EWS/ETS was seen to function as a co-activator for TERT transcription. A number of oncogenes and cancer-related genes contain AU-rich element (ARE) in non-coding regions of transcribed mRNA. HuR is a RNA-binding protein that has the potential to stabilize ARE-containing mRNAs. HuR is known to shuttle between the nucleus and the cytoplasm via several export pathways. When normal cells are exposed to stress, HuR is exported to the cytoplasm in a chromosome maintenance region 1 (CRM1)-dependent manner. However, we demonstrate that HuR is CRM-1 independently exported to the cytoplasm in oral cancer cells. ARE-mRNAs were also exported to the cytoplasm and stabilized in the oral cancer cells, which were inhibited by HuR knockdown. These findings suggest that transcriptional and translational abnormalities of oncogenes may contribute to the carcinogenesis of oral epithelial cells. © 2009, Japanese Stomatological Society. All rights reserved.
• Attenuated p53 activation in tumour-associated stromal cells accompanies decreased sensitivity to etoposide and vincristine

  A. C. Dudley, S-C Shih, A. R. Cliffe, K. Hida, M. Klagsbrun

  BRITISH JOURNAL OF CANCER 99 (1) 118 - 125 0007-0920 2008/07 [Refereed][Not invited]
   

  Alterations in the tumour suppressor p53 have been reported in tumour-associated stromal cells; however, the consequence of these alterations has not been elucidated. We investigated p53 status and responses to p53-activating drugs using tumour-associated stromal cells from A375 melanoma and PC3 prostate carcinoma xenografts, and a spontaneous prostate tumour model ( TRAMP). p53 accumulation after treatment with different p53-activating drugs was diminished in tumour-associated stromal cells compared to normal stromal cells. Tumour-associated stromal cells were also less sensitive to p53-activating drugs - this effect could be reproduced in normal stromal cells by p53 knockdown. Unlike normal stromal cells, tumour stromal cells failed to arrest in G(2) after etoposide treatment, failed to upregulate p53-inducible genes, and failed to undergo apoptosis after treatment with vincristine. The lower levels of p53 in tumour stromal cells accompanied abnormal karyotypes and multiple centrosomes. Impaired p53 function in tumour stroma might be related to genomic instability and could enable stromal cell survival in the destabilising tumour microenvironment.
• Expression of E1AF, an ets-oncogene transcription factor, highly correlates with malignant phenotype of malignant melanoma through up-regulation of the membrane-type-1 matrix metalloproteinase gene

  Hironobu Hata, Tetsuya Kitamura, Fumihiro Higashino, Kyoko Hida, Koichi Yoshida, Yoichi Ohiro, Yasunori Totsuka, Yoshimasa Kitagawa, Masanobu Shindoh

  ONCOLOGY REPORTS 19 (5) 1093 - 1098 1021-335X 2008/05 [Refereed][Not invited]
   

  Matrix metalloproteinase (MMP) is closely involved in the degradation of extracellular matrix and confers invasive and metastatic potential to malignant tumors. MMP-2 is a type-IV collagenase secreted as a proenzyme that is activated on the surface of the tumor cell by membrane-type I MMP (MT1-MMP). MT1-MMP plays a critical role during tumor progression and metastasis. We investigated the expression levels of E1AF and MT1-MMP in malignant melanoma cell lines and specimens from patients in order to clarify the mechanisms responsible for the invasion and metastasis of malignant melanoma. High levels of E1AF and MT1-MMP mRNA expression were observed in melanoma cells by Northern blotting and real-time PCR. The expression level was highly correlated with an invasive potential determined by an in vitro invasion assay. The down-regulation of MT1-MMP was identified when E1AF was knocked down by RNA interference. These results suggest that E1AF plays a crucial role in the invasion and metastasis of malignant melanoma through up-regulating the MT1-MMP expression.
• Understanding tumor endothelial cell abnormalities to develop ideal anti-angiogenic therapies

  Kyoko Hida, Yasuhiro Hida, Masanobu Shindoh

  CANCER SCIENCE 99 (3) 459 - 466 1349-7006 2008/03 [Refereed][Not invited]
   

  Tumor angiogenesis is necessary for solid tumor progression and metastasis. Tumor blood vessels have been shown to differ from their normal counterparts, for example, by changes in morphology. An important concept in tumor angiogenesis is that tumor endothelial cells are assumed to be genetically normal, even though these endothelial cells are structurally and functionally abnormal. To date, many anti-angiogenic drugs have been developed, but, their therapeutic efficacy is not dramatic and they have also been reported to cause toxic side effects. To develop ideal antiangiogenic therapies, understanding tumor endothelial cell abnormalities is important. We have isolated tumor endothelial cells from mouse tumor xenografts and have shown that tumor-associated endothelial cells are abnormal. Tumor-associated endothelial cells upregulate many genes, such as epidermal growth factor receptor (EGFR). Tumor-associated endothelial cells are also more sensitive to EGF. They also have relatively large, heterogeneous nuclei. Unexpectedly, tumor endothelial cells are cytogenetically abnormal. Fluorescence in situ hybridization (FISH) analysis showed that freshly isolated uncultured tumor endothelial cells were aneuploid and had abnormal multiple centrosomes. The degree of aneuploidy was exacerbated by passage in culture. In marked contrast, freshly isolated normal skin and adipose endothelial cells were diploid. They had normal centrosomes and remained cytogenetically stable in culture even up to 20 passages. We conclude that tumor endothelial cells can acquire cytogenetic abnormalities while in the tumor microenvironment. Questions as to whether or not tumor endothelial cells become resistant to antiangiogenic drugs are thus raised. Our preliminary data show that tumor endothelial cells are more resistant to certain chemotherapeutic drugs. Studies to evaluate the mechanism for cytogenetic abnormalities in tumor endothelial cells are underway. It is becoming quite clear that the tumor vasculature is much more complex and unpredictable than initially perceived. Here, we provide an overview of the current studies on tumor endothelial cell abnormalities.
• OP-193 Adrenomedullin antagonist 発現ベクターを用いたヒト腎癌移植マウスモデルに対する遺伝子治療(第95回日本泌尿器科学会総会)

  土屋 邦彦, 樋田 京子, 村木 力, 近藤 健, 三関 哲矢, 進藤 正信, 原林 透, 篠原 信雄, 小林 正伸, 野々村 克也

  日本泌尿器科学会雑誌 一般社団法人 日本泌尿器科学会 98 (2) 339 - 339 2007
• Tumor endothelial cells express epidermal growth factor receptor (EGFR) but not ErbB3 and are responsive to EGF and to EGFR kinase inhibitors

  DN Amin, K Hida, DR Bielenberg, M Klagsbrun

  CANCER RESEARCH 66 (4) 2173 - 2180 0008-5472 2006/02 [Refereed][Not invited]
   

  Epidermal growth factor (EGF) receptor family members are expressed by tumor cells and contribute to tumor progression. The expression and activity of EGF receptors in endothelial cells are less well characterized. Analysis of tumor-derived endothelial cells showed that they express EGFR, ErbB2, and ErbB4, whereas their normal counterparts express ErbB2, ErbB3, and ErbB4. The gain in expression of EGFR and the loss of ErbB3 expression in tumor vasculature was also observed in vivo. As a consequence of their expressing EGFR, tumor endothelial cells responded to EGF and other EGF family members by activating both EGFR and ErbB2, by activating the downstream mitogen-activated protein kinase pathway, and by enhanced proliferation. On the other hand, normal endothelial cells did not respond to EGF but instead were responsive to neuregulin (NRG), a ligand for ErbB3 and ErbB4. NRG activated ErbB3 in normal endothelial cells and inhibited growth of these cells. In contrast, tumor endothelial cells, which do not express ErbB3, were not growth inhibited by NRG. Furthermore, due to their expression of EGFR, tumor endothelial cells, unlike normal endothelial cells, are direct targets for EGFR kinase inhibitors. These low-molecular-weight compounds block EGF-induced EGFR activation and proliferation of tumor endothelial cells. These results suggest that a gain of EGF-induced endothelial cell proliferation, and loss of NRG-induced growth inhibition in tumor endothelial cells constitutes a switch that promotes tumor angiogenesis. In addition, these results suggest that EGFR kinase inhibitors may be effective for antiangiogenesis therapy by specifically targeting the tumor, but not the normal, vasculature.
• A new perspective on tumor endothelial cells: Unexpected chromosome and centrosome abnormalities

  K Hida, N Klagsbrun

  CANCER RESEARCH 65 (7) 2507 - 2510 0008-5472 2005/04 [Refereed][Not invited]
   

  it has been shown that endothelial cells in solid tumors are cytogenetically abnormal. These cells are aneuploid with multiple chromosomes and multiple centrosomes. Unlike normal endothetial cells which remain diploid in long-term culture, the aneuploidy of tumor endothelial cells is exacerbated in culture suggesting that these cells are inherently unstable. It is speculated that this instability might compromise the effectiveness of antiangiogenesis therapy.
• Tumor-associated endothelial cells with cytogenetic abnormalities

  K Hida, Y Hida, DN Amin, AF Flint, D Panigrahy, CC Morton, M Klagsbrun

  CANCER RESEARCH 64 (22) 8249 - 8255 0008-5472 2004/11 [Refereed][Not invited]
   

  Tumor angiogenesis is necessary for solid tumor progression and metastasis. Tumor blood vessels have been shown to differ from normal counterparts, for example, by changes in morphology. An important concept in tumor angiogenesis is that tumor endothelial cells are assumed to be genetically normal, although these endothelial cells are structurally and functionally abnormal. However, we hypothesized that given the phenotypic differences between tumor and normal blood vessels, there may be genotypic alterations as well: Mouse endothelial cells were isolated from two different human tumor xenografts, melanoma and liposarcoma, and from two normal endothelial. cell counterparts, skin and adipose. Tumor-associated endothelial cells expressed typical endothelial cell markers, such as CD31. They had relatively large, heterogeneous nuclei. Unexpectedly, tumor endothelial cells were cytogenetically abnormal. Fluorescence in situ hybridization (FISH) analysis showed that freshly isolated uncultured tumor endothelial cells were aneuploid and had abnormal multiple centrosomes. The degree of aneuploidy was exacerbated by passage in culture. Multicolor FISH indicated that the structural chromosomal aberrations in tumor endothelial cells were heterogeneous, indicating that the cytogenetic alterations were not clonal. There was no evidence of human tumor-derived chromosomal material in the mouse tumor endothelial cells. In marked contrast, freshly isolated normal skin and adipose endothelial cells were diploid, had normal centrosomes, and remained cytogenetically stable in culture even up to 20 passages. FISH analysis of tumor sections also showed endothelial cell aneuploidy. We conclude that tumor endothelial cells can acquire cytogenetic abnormalities while in the tumor microenvironment.
• Arthrocentesis following arthroscopic examination in patients with closed lock of the temporomandibular joint

  YURA Shinya, TOTSUKA Yasunori, OOI Kazuhiro, MABUCHI Akiko, HORIMUKAI Hiromasa, ASHIKAGA Yuichi, DEYAMA Ayako, YOSHIKAWA Tetsuya, OHIRO Yoichi, HIDA Kyoko

  Journal of Oral Surgery Society of Japan Japanese Society of Oral and Maxillofacial Surgeons 49 (4) 246 - 251 0021-5163 2003 [Refereed][Not invited]
   

  We describe procedures for arthrocentesis based on arthroscopic findings and report the therapeutic usefulness of these procedures in 46 patients with closed lock of the temporomandibular joint.
  Arthroscopic examination was done under local anesthesia in the outpatient clinic. An 18-gauge needle and a trocar for a 0.8-mm rod-lens fiberscope were inserted into the anterior and posterior compartments of the upper joint space, respectively. The anterior compartment was then observed with special respect to the presence of adhesion. In joints without adhesion, irrigation was done under low hydraulic pressure. In joints with adhesion, irrigation was performed under sufficient hydraulic pressure (maximum pressure exerted, 40 KPa) to widen the upper joint space. Manipulation was performed after irrigation.
  Eight weeks after these two procedures, the range of maximum mouth opening increased from 2 to 20 mm and averaged 10.3 mm in 18 patients without adhesion. In 28 patients with adhesion, the increase in maximum mouth opening ranged 3 to 25 mm and averaged 9.1 mm. There was no significant difference in the increase in maximum mouth opening between these two groups.
• Role of neuropilins and semaphorins in angiogenesis and cancer

  M Kreuter, D Bielenberg, Y Hida, K Hida, M Klagsbrun

  ANNALS OF HEMATOLOGY 81 S74 - S74 0939-5555 2002 [Refereed][Not invited]
  
• BAG-1 expression correlates highly with the malignant potential in early lesions (T1 and T2) of oral squamous cell carcinoma

  M. Shindoh, M. Adachi, F. Higashino, M. Yasuda, K. Hida, T. Nishioka, M. Ono, S. Takayama, J.C Reed, K. Imai, Y. Totsuka, T. Kohgo

  Oral Oncology 36 (5) 444 - 449 1368-8375 2000/09 [Refereed][Not invited]
   

  BAG-1 is a Bcl-2-binding protein that functions as an anti-apoptotic molecule. In this report we show a possible correlation between BAG-1 expression levels and the probability of oral squamous cell carcinoma (SCC) progression. We investigated BAG-1 expression levels in 22 patients diagnosed with early lesions (T1 and T2) of oral SCCs using immunohistochemistry and western blotting. High steady-state levels of BAG-1 were detected in 13 out of 22 cases (59%). High BAG-1 expression was observed more frequently in cases with nodal metastasis (89%) than in those without nodal metastasis (38%) (P < 0.03), suggesting that BAG-1 expression levels may correlate with the pathological stage of oral SCCs. Furthermore, BAG-1 expression levels correlated with the WHO grade, i.e. 45% in grade-I cases as opposed to 72% in grade-II cases (P < 0.02). These data suggest that an analysis of BAG-1 expression may be useful in establishing a prognosis for patients with oral SCCs, and especially in predicting the metastatic potential of SSCs. (C) Elsevier Science Ltd.
• Hepatocyte growth factor upregulates E1AF that induces oral squamous cell carcinoma cell invasion by activating matrix metalloproteinase genes

  M Hanzawa, M Shindoh, F Higashino, M Yasuda, N Inoue, K Hida, M Ono, T Kohgo, M Nakamura, K Notani, H Fukuda, Y Totsuka, K Yoshida, K Fujinaga

  CARCINOGENESIS 21 (6) 1079 - 1085 0143-3334 2000/06 [Refereed][Not invited]
   

  Hepatocyte growth factor (HGF) is thought to play a role in cell motility and invasion. Matrix metalloproteinases (MMPs) have been implicated in invasion and metastasis of tumor cells. We have previously reported that the Ets-oncogene family transcription factor E1AF positively regulates transcription of MMP genes in transient expression assays and that overexpression of the E1AF gene confers an invasive phenotype on breast cancer cells. Here we examined the effect of HGF on E1AF and MMP gene expression in terms of the invasive potential of the oral squamous cell carcinoma cell line HSC3. HGF stimulated expression of the E1AF gene. The levels of MMP-1, -3 and -9 mRNAs increased in cells treated with HGF and correlated with E1AF upregulation, In contrast, no obvious upregulation of MMP-1 and -9 mRNA was observed in ASE1AFHSC3 cells transfected with the antisense E1AF expression vector into parental HSC3 cells. The wild-type MMP-9 gene promoter was activated by endogenous E1AF in HSC3 cells, and chloramphenicol acetyltransferase (CAT) activities increased when HGF was added to transfected cells. On the other hand, CAT activity was reduced to almost two-thirds of the wild-type activity when HSC3 cells were transfected with a CAT reporter plasmid driven by a mutant MMP-9 promoter lacking the Ets-binding site, and induction of CAT activity was not observed upon addition of HGF. Analysis of organotypic raft cultures revealed that HSC3 cells invaded and degraded collagen gel actively upon addition of HGF. These results suggest that HGF induces expression of the Ets-related E1AF transcription factor gene whose product in turn activates MMP genes and leads to oral cancer cell invasion.
• Practical application of HPC film as an intraoral covering material

  K. Kasashi, S. Harada, I. Wada, I. Kobayashi, K. Funaoka, K. Hida, Y. Totsuka

  Oral Therapeutics and Pharmacology 18 (3) 110 - 115 0288-1012 1999 [Refereed][Not invited]
   

  When administering medication, the oral cavity presents specific problems in comparison with other organs. In other words, there are diverse factors such as the palate, the proximity to olfaction, the existence of sensitive tactus and the tongue, sputum and teeth. Carboxymethyl Cellulose Sodium (CMC) and Sodium Polyacrylate have been used as adhesive agents for the mucous membrane of the oral cavity in manufactured oral ointments and hospital preparations. However, even with the use of these agents, the adhesion time of oral ointments has not increased satisfactorily and the problem of the taste of the ingredients remains. This situation requires improvements for comfortable use of such ointments for all occasions. HPC Film consisting of Hydroxypropyl Cellulose (HPC), a water-soluble high- polymer was used as a covering material without active ingredients. After applying 10% Tetracycline CMC ointment to the buccal mucosa, lower buccal gingiva, soft palate and margin of tongue, they were covered by HPC Film. Changes over time in the presence of the ointment were compared between cases with and without HPC Film. It was found that by using a covering of HPC Film, adhesion time was markedly increased. Even in the tongue which was difficult to attach, the adhesion-maintenance time of oral ointment with HPC film indicated six-fold that without the film.
• Activation of the p21(Waf1/Cip1) promoter by the ets oncogene family transcription factor E1AF

  K Funaoka, M Shindoh, K Yoshida, M Hanzawa, K Hida, S Nishikata, Y Totsuka, K Fujinaga

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 236 (1) 79 - 82 0006-291X 1997/07 [Refereed][Not invited]
   

  p21(Waf1/Cip1) is one of the key regulatory proteins in cell cycle, terminal differentiation, and apoptosis. Its promoter was shown to be transactivated by the wild-type p53 protein as well as in a p53-independent manner. In this report, we demonstrate that E1AF, an ets-related transcription factor, activated the human p21(Waf1/Cip1) promoter by interacting with the ets-binding sites located close to the two previously identified p53-responsive elements. Northern blot analysis revealed that p21(Waf1/Cip1) and E1AF were correlatively upregulated in response to cisplatin treatment in SiHa cells. Transient expression assays demonstrated that E1AF can activate the p21(Waf1/Cip1) promoter-driven luciferase reporter gene in SiHa cells. The p21(Waf1/Cip1) promoter activity was also increased in p53-null Saos2 osteosarcoma cells, but was markedly reduced when the ets-binding sites were deleted. These results indicate that E1AF positively regulates transcription from the p21(Waf1/Cip1) promoter in response to genotoxic stresses. (C) 1997 Academic Press.
• Antisense E1AF transfection restrains oral cancer invasion by reducing matrix metalloproteinase activities

  K Hida, M Shindoh, M Yasuda, M Hanzawa, K Funaoka, T Kohgo, A Amemiya, Y Totsuka, K Yoshida, K Fujinaga

  AMERICAN JOURNAL OF PATHOLOGY 150 (6) 2125 - 2132 0002-9440 1997/06 [Refereed][Not invited]
   

  E1AF is a newly identified human ets-family transcription factor We have reported that E1AF can up-regulate transcription of matrix metalloproteinase (MMP) genes and confers invasive phenotype on human cancer cells. HSC3 is an oral squamous-cell-carcinoma-derived cell line, and it manifests high levels of E1AF and MMP-1 and -9 gene expression that are associated with invasive potential, We reconstructed an E1AF antisense expression vector, transfected HSC3 cells with the vector, and obtained HSC3AS cells that express E1AF antisense RNA, HSC3AS showed decreasing mRNA and protein levels of MMP-1, -3, and -9, Moreover, HSC3AS showed lower invasive potential in vitro three-dimensional raft culture and in vivo implantation into nude mice. These results imply that transfection of antisense E1AF inhibits tumor invasion by down-regulating MMP genes.
• Expression of E1AF, an ets-family transcription factor, is correlated with the invasive phenotype of oral squamous cell carcinoma

  K. Hida, M. Shindoh, K. Yoshida, A. Kudoh, K. Furaoka, T. Kohgo, K. Fujinaga, Y. Totsuka

  Oral Oncology 33 (6) 426 - 430 1368-8375 1997 [Refereed][Not invited]
   

  E1AF is a newly identified ets-oncogene family transcription factor. Previous reports have noted that E1AF can upregulate promoter activities of several matrix metalloproteinase (MMP) genes and showed that invasive potentials of oral squamous cell carcinoma-derived cell lines are correlated with expression of E1AF and MMPs. The invasive phenotype is restrained by transfection with an antisense E1AF expression vector. Thus, E1AF is thought to be highly correlated with malignant potentials of cancer cells. However, little is known about E1AF expression and cancer cell malignancies in in vivo tumours. In the present study, 27 oral squamous cell carcinoma (SCC) specimens were examined using RT-PCR, Southern blot hybridisation and in situ hybridisation (ISH) and compared to the clinicopathological parameters. Among the 27 patients, E1AF was detected in 15 cases. E1AF mRNA was detected in 13 of 17 invasive SCCs, whereas the majority of SCCs not expressing E1AF showed an expansive growth pattern. Increased prevalence of E1AF-positive oral SCC was observed in cases with nodal metastasis. These results indicate that E1AF may be involved in cancer cell malignancies through its ability to promote invasive potential.
• E1AFアンチセンス発現ベクター導入による口腔扁平上皮癌細胞の浸潤能の抑制およびE1AF発現の臨床的意義

  Kyoko Hida

  北海道歯学雑誌 18 (2) 105 - 114 1997 [Refereed][Not invited]
  
• [PCR application for paraffin-embedded tissues].

  Shindoh M, Fujinaga K, Hida K, Funaoka K

  Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 5 41 463 - 466 0039-9450 1996/04 [Not refereed][Invited]
  
• Correlated expression of matrix metalloproteinases and ets family transcription factor E1A-F in invasive oral squamous-cell-carcinoma-derived cell lines

  M Shindoh, F Higashino, M Kaya, M Yasuda, K Funaoka, M Hanzawa, K Hida, T Kohgo, A Amemiya, K Yoshida, K Fujinaga

  AMERICAN JOURNAL OF PATHOLOGY 148 (3) 693 - 700 0002-9440 1996/03 [Refereed][Not invited]
   

  Matrix metalloproteinases (MMPs) have been implicated irt invasion and metastasis of tumor cells. Transcription regulatory regions of MMP genes often contain binding sites for ets transcription factors. We recently isolated a cDNA encoding human E1A-F, a member of the ets oncogene family, and showed that E1A-F; can upregulate MMP genes by CAT assay. We attempted to investigate the relationship between E1A-F mRNA expression and MMP protein expression in four different types of oral squamous-cell-carcinoma-derived cell lines (HSC 3, SAS, KB, and Ca 9.22). HSC 3 and SAS are highly invasive cell lines when they are injected in the tongue of nude mice Raft culture of HSC 3 and SRS revealed the same characteristics as seen in tumors implanted in viva Both type I collagenase (MMP-1) and 92-kd type IV collagenase (MMP-9) were detected in cultured HSC 3 and SAS cells. E1A-F mRNA was demonstrated to be highly expressed in HSC 3 and SAS by Northern blotting, and in situ hybridization confirmed E1A-F mRNA expression at the invasion front of tumor cells seeded on collagen gel. On the other hand, RE and Ca 9.22 have little potential for invasion, and MMP-1 and MMP-9 protein and E1A-F mRNA could not be detected. These results suggest that the ets-related E1A-F participates in the regulation of invasion-associated MMP genes and is involved ia presenting invasive activity in tumor cells of oral squamous cell carcinoma.
• Synergistic enhancement of cellular uptake with CD44-expressing malignant pleural mesothelioma by combining cationic liposome and hyaluronic acid-lipid conjugate

  Sakurai Y, Kato A, Hida Y, Hamada J, Maishi N, Hida K, Harashima H

  J Pharm Sci [Refereed][Not invited]
  
• Tumor endothelial heterogeneity in cancer progression

  Nako Maishi, Dorcas A. Annan, Hiroshi Kikuchi, Yasuhiro Hida, Kyoko Hida

  Cancers [Refereed][Not invited]
  

Books etc

• 新臨床腫瘍学（ 改訂第7版）ーがんの発生と進展機構ー

  樋田京子 (Contributor血管新生 P49-52)
  南江堂 2024/02
• 新臨床腫瘍学（改訂第6版）－がん薬物療法専門医のために－

  樋田京子 (Contributor血管新生 P42-46)
  南江堂 2021/05
• 高倉伸幸企画, がん免疫の効果を左右する腫瘍血管と免疫環境

  樋田京子, 間石奈湖 (Contributor腫瘍血管内皮細胞の異常性と腫瘍免疫, 3201-3205,)
  羊土社 2019/11
• Cancer Drug Delivery Systems Based on the Tumor Microenvironment.

  Hida K, Maishi N, Hida Y (ContributorTumor Blood Vessels as Targets for Cancer Therapy. p41-56)
  Springer, Tokyo, 2019
• 新臨床腫瘍学（改訂第5版）－がん薬物療法専門医のために－

  樋田京子 (Contributor血管新生 P48-51)
  南江堂 2018/07
• がん創薬フロンティア Part Ⅴ

  樋田京子 (Contributor第19章 血管新生阻害剤)
  化学同人編 2018
• パラダイムシフトをもたらすエクソソーム機能研究最前線

  樋田京子, 間石奈湖, 森本真弘 (Contributorがんとエクソソーム)
  エヌ・ティー・エス 2017
• Cancer Metastasis and Cancer Stem Cell/Niche, “Tumor Endthelial Cells and Cancer Progression

  Hida K, Maishi N, Dorcas Akuba-Muhyia Annan, Hida Y (ContributorTumor Endthelial Cells and Cancer Progression”)
  Bentham Books 2016
• 別冊BIO Clinica 慢性炎症と疾患～慢性炎症とがん

  樋田京子, 大賀則孝, 間石奈湖, 秋山廣輔, 樋田泰浩 (Contributor腫瘍血管内皮細胞の多様性)
  北隆館 2016/01
• 口腔組織・発生学 第2版

  樋田 京子 (Contributor癌と血管新生)
  医歯薬出版 2015/02
• 新臨床腫瘍学（改訂第4版）－がん薬物療法専門医のために－

  樋田 京子 (Contributor血管新生)
  南江堂 2015
• 血管新生研究の最先端

  HIDA Kyoko (Contributor血管新生と病態－腫瘍血管新生)
  2013
• 血管生物医学事典

  樋田 京子 (Contributor腫瘍血管内皮細胞)
  朝倉書店 2011
• がん転移研究の実験手法 第1版

  樋田 京子 (Contributor磁気ビーズを用いたがん組織からの細胞分離法)
  金芳堂 2008

Conference Activities & Talks

• Tumor endothelial cells and inflammaging  [Invited]

  Kyoko Hida

  第113回日本病理学会総会, 先端的分子病理学講習会 分子腫瘍病理学の新展開  2024/03
• 血管から考えるがん転移とCOVID19重症化の制御  [Invited]

  Kyoko Hida

  藤田医科大学 2023年度第9回医学講演  2024/03
• Contribution of tumor endothelial cells in tumor progression  [Invited]

  Hida K

  The 1st International Symposium on Cancer Immunology and Immunotherapy  2024/03
• 血管研究の先に見えるもの  [Invited]

  樋田京子

  第 35 回三省堂サイエンスカフェ in 札幌 北海道大学CoSTEP 日本学術会議北海道地区会議 共催  2024/03
• 血管病態研究からがん臨床への展開の試み”, 2024.1.26（札幌）  [Invited]

  樋田京子

  第42回日本口腔腫瘍学会総会・学術大会特設講演  2024/01
• 腫瘍血管の異常性とがんの悪性化  [Invited]

  樋田京子

  京都大学医生物学研究所 再生組織構築研究部門 生体材料学分野セミナー  2024/01
• 腫瘍血管の炎症老化

  樋田京子

  金沢大学がん進展制御研究所セミナー  2023/11
• 腫瘍血管内皮細胞による血栓形成促進  [Invited]

  樋田京子

  第64回日本脈管学会学術総会シンポジウム  2023/10
• Acquisition of endthelial cell abnormality by highly metastatic tumor EV  [Invited]

  森本真弘, 間石奈湖, 北川善政, 樋田京子

  第10回日本細胞外小胞学会学術集会  2023/10
• The oral bacterium Streptococcus mutans promotes tumor metastasis by inducing vascular inflammaƟon and thrombosis  [Invited]

  Yu L, Nako Maishi N, Akahori E, Hong Y, Hasebe A, Takeda R, Matsuda A, Hida Y, Kitagawa Y, Hida K

  Vascular Biology 2023  2023/10
• 腫瘍血管内皮細胞とがん微小環境”, 2023.10.7  [Invited]

  樋田京子

  第88回日本泌尿器科学会 東部総会 札幌 特別講演  2023/10
• Targeting cancer and vascular crosstalk for cancer therapy

  Maishi N, Hida K

  The 82nd annual meeting of the Japanese Cancer Association International Sessions “New horizons in tumor microenvironment biology for cancer therapy  2023/09
• The oral bacterium Streptococcus mutans promotes tumor metastasis by inducing vascular inflammation  [Invited]

  Li Yu, Maishi N, Akahori E, Hasebe A, Takeda R, Yanagawa Matsuda A, Hida Y, Nam JM, Onodera Y, Kitagawa Y, Hida K

  第82回日本癌学会学術総会 [YSA]Cancer Science ヤングサイエンティストアワード受賞講演  2023/09
• 腫瘍血管内被細胞によるがん転移促進  [Invited]

  樋田京子

  第65回歯科基礎医学会学術大会アップデートシンポジウム  2023/09
• Analysis of lung endotheliopathy in lethal mice infected with mouse- adapted SARS-CoV-2  [Not invited]

  Hida K

  2023 KVBM Annual Meeting & AAVBM in partnership with ICoLA  2023/09
• 血管の多様性とがんの悪性化  [Invited]

  樋田京子

  第41回日本ヒト細胞学会学術集会シンポジウム  2023/08
• がん, 感染症における血管―微小環境ネットワーク”, 2023.7.27（東京）  [Invited]

  樋田京子

  第41回日本骨代謝学会学術集会シンポジウム  2023/07
• がん細胞－血管内皮細胞Cluster形成によるがん悪性化促進  [Invited]

  間石奈湖, 樋田京子

  第32回日本がん転移学会学術集会・総会  2023/07
• 癌微小環境と血管内皮細胞のネットワーク”  [Invited]

  樋田京子

  第43回日本骨形態計測学会  2023/06
• Vasclar endothelial cell heterogeneity in diseases  [Invited]

  Hida K

  The 2023 Conference of Asia Pacific Societies for Extracellular Vesicles（APSEV）  2023/04
• 腫瘍血管内皮細胞の多様性と治療への応用  [Invited]

  樋田京子

  北海道DDS研究会 公開特別シンポジウム  2023/03
• 血管の多様性～Wetにdryを取り入れて  [Invited]

  樋田京子

  北海道発産官学・地域連携ダイバーシティ 数理・データサイエンス 特別セッション・ワークショップ  2023/02
• 血管とがんの悪性化  [Invited]

  樋田京子

  日本女性科学者の会新春シンポジウム2023 学術講演会  2023/01
• ，“腫瘍血管の多様性とがんの悪性化”，2022.12.13（札幌）  [Invited]

  樋田京子

  AIR－Angiogenesis & Immunology Research－  2022/12
• The heterogeneity of endothelial cells in tumor and infectious disease  [Invited]

  樋田京子

  第45回日本分子生物学会年会公募ワークショップ「生体組織の形成・再生・恒常性維持とその破綻における血管の新たな機能」  2022/12
• Tumor endothelial cells induce metastasis by disrupting immune environment  [Invited]

  Hida K

  22nd International Vascular Biology Meeting (IVBM 2022)  2022/10
• 腫瘍血管とがん微小環境  [Invited]

  Hida K

  Gynecologic Cancer Clinical Research Seminar 2022  2022/10
• Epigenetic heterogeneity in tumor endothelial cells  [Invited]

  Hida K

  The 81st Annual Meeting of the Japanese Cancer Association  2022/10
• The contribution of tumor endothelial cells in tumor progression  [Invited]

  Hida K

  The 81st Annual Meeting of the Japanese Cancer Association, JCA-AACR Joint Symposium -Sponsored by Princess Takamatsu Cancer Research Fund-  2022/09
• 腫瘍血管内被細胞の特性とがん免疫（誌上発表）  [Invited]

  樋田京子

  第64回歯科基礎医学会学術大会 アップデートシンポジウム5「4歯科基礎領域におけるがん研究フロンティア」  2022/09
• 双子連れ留学体験記～ハンディがあってもなんとかなる！？,2022.9,12（WEB開催）  [Invited]

  樋田京子

  一般社団法人 日本歯科医学会連合 主催 令和４（2022)年度 国際活動委員会フォーラム“そうだ留学、しよう！～逆転の発想！ 日本人だからこそ留学しよう～”  2022/09
• Contribution of tumor endotheilal cells in tumor progression  [Invited]

  Hida K

  The 7th JCA－AACR Special Joint Conference“The Latest Advances in Pancreatic Cancer Research:From Basic Science to Therapeutics”  2022/07
• The impact of vascular endothelial cell heterogeneity in tumor microenvironment  [Invited]

  Hida K

  The 40th Sapporo International Cancer Symposium  2022/06
• 腫瘍血管内皮細胞から癌免疫微小環境への作用  [Invited]

  樋田京子

  第2回血管研究会  2022/02
• 腫瘍血管とがん微小環境  [Invited]

  Kyoko Hida

  KCOG Scientific Seminar  2022/01
• 腫瘍血管とがん微小環境のクロストーク

  Kyoko Hida

  CABOMETYX Round TableMeeting in East Japan  2022/01
• 女性活躍への展望〜北海道大学歯学部における男女共同参画の現状から〜  [Invited]

  樋田京子

  日本学術会議歯学委員会, 病態系歯学分科会主催, 一般財団法人日本歯科医学会連合共催公開シンポジウム“歯学分野におけるジェンダー・ダイバーシティ～課題と展望について～”  2022/01
• 腫瘍血管内皮細胞とがん免疫微小環境  [Invited]

  樋田京子

  第2回Oncology Crosstalk Symposium  2021/12
• 腫瘍血管とがん微小環境  [Invited]

  樋田京子

  鳥取大学医学部セミナー  2021/11
• The role of tumor endothelial cells in cancer metastasis  [Invited]

  Kyoko Hida

  The 80nd Annual Meeting of the Japanese Cancer Association  2021/09
• 免疫療法に先行した血管新生阻害剤投与は治療効果を改善する

  佐藤 峰嘉, 間石 奈湖, 樋田 泰浩, 松田 彩, 今野 哲, 樋田 京子

  日本癌学会総会記事  2021/09  (一社)日本癌学会
  
• Biglycanを標的とした新規血管新生阻害剤候補薬の検討

  竹川 英輝, 間石 奈湖, 松田 彩, 樋田 泰浩, 樋田 京子

  日本癌学会総会記事  2021/09  (一社)日本癌学会
  
• 腫瘍血管内皮細胞による微小環境の乱れとがんの転移  [Invited]

  樋田京子

  第30回日本がん転移学会学術集会・総会シンポジウム「血管から転移を考える」  2021/07
• 口腔扁平上皮癌における高内皮細静脈と臨床的因子との関連

  新山 宗, 間石 奈湖, 松田 彩, 鄭 漢忠, 大廣 洋一, 上田 倫弘, 樋田 京子

  日本口腔科学会雑誌  2021/07  (NPO)日本口腔科学会
  
• 腫瘍血管制御によるがん免疫活性化  [Invited]

  Kyoko Hida

  第4回AIR in Sapporo－Angiogenesis & Immunology Research－  2021/06
• 腫瘍血管内皮細胞の特性とがん免疫  [Invited]

  樋田京子

  第110回日本病理学会総会シンポジウム「がんと免疫病理, 最前線」  2021/04
• The Role of tumor endothelial cells in cancer progression  [Invited]

  Kyoko Hida

  The 94th Annual Meeting og the Japanese Pharmacological Society  2021/03
• “私のWork-Life Blend”, 2021.3.1（WEB開催）  [Invited]

  樋田京子

  北海道大学病院男女共同参画推進室・北海道大学大学院歯学研究院FD委員会共催 男女共同参画推進室講演会「キャリアを考える」  2021/03
• 口腔癌における高内皮細静脈の役割の検討

  松田 彩, 新山 宗, 間石 奈湖, 上田 倫弘, 樋田 京子

  日本病理学会会誌  2021/03  (一社)日本病理学会
  
• FGF2と新規ケイ酸カルシウム系材料を用いた生活歯髄切断法の基礎的研究

  近 加名代, 北村 哲也, 松田 彩, 間石 奈湖, Islam Rafiqul, 戸井田 侑, 佐野 英彦, 樋田 京子

  北海道歯学雑誌  2021/03  北海道歯学会
   

  生活歯髄切断法は、冠部歯髄を除去し根部歯髄を保存する方法で、通常乳歯や幼若永久歯に用いられる。歯根完成後の永久歯の歯髄に不可逆的な炎症が生じると、抜髄処置の適応となることが多い。乳歯や幼若永久歯は、歯髄細胞の増殖活性が高く、血液循環が良好であるため生活歯髄切断法の成功率が高いと考えられている。しかし、歯根完成後は歯髄の修復に重要な血流量の減少や、歯髄幹細胞の存在が期待できないことから、生活歯髄切断法による治療は困難であると考えられる。したがって、歯髄修復作用のある材料を用いた生活歯髄切断法の開発は永久歯の歯髄保存療法における重要な課題となっている。そこで本研究では、塩基性線維芽細胞増殖因子(FGF2)と新規ケイ酸カルシウム系材料(WPPT)を覆髄材として使用し、ラットの生活歯髄切断後の歯髄修復に及ぼす作用を組織学的に検討した。生活歯髄切断後の歯髄の炎症性変化を経時的に観察したところ、7日目ではFGF2とWPPTを併用した群で高度な好中球浸潤がみられたが、他の群ではほとんど好中球浸潤はみられなかった。30日目では、FGF2とWPPTを併用した群では炎症性変化が認められなかったが、他の群では歯髄壊死や好中球浸潤がみられた。形成されたデンティンブリッジの面積は、7日目、14日目ではすべての群で同程度であったが、30日目ではFGF2を含む群で他の群と比較して大きく、FGF2がデンティンブリッジの形成を促進した可能性が考えられた。残存歯髄の血管について評価したところ、FGF2とWPPTを併用した群では7日目で血管新生がみられ、30日目で歯髄の炎症が消退したことが示唆された。他の群では30日目にも炎症が持続していることが示唆された。以上の結果より、FGF2とWPPTの併用は、残存歯髄の正常な修復を促進し、歯髄保存を可能とすることが示唆された。本研究により、FGF2とWPPTを併用することで、歯根完成後の永久歯にも生活歯髄切断法を応用できる可能性が示唆された。(著者抄録)
• 泌尿器癌における腫瘍血管内皮細胞の役割

  菊地 央, 山田 修平, 古御堂 純, 宮田 遥, 松本 隆児, 大澤 崇宏, 秋野 文臣, 土屋 邦彦, 安部 崇重, 間石 奈湖, 樋田 京子, 篠原 信雄

  日本泌尿器科学会総会  2020/12  (一社)日本泌尿器科学会総会事務局
  
• 腫瘍血管と免疫小環境  [Invited]

  Kyoko Hida

  Breast Cancer Investigator’s Meeting in Sapporo,  2020/11
• Co-existing with SARS-CoV-2 新型コロナウイルス時代の提案 口腔からのモデリングアプローチによる感染対策

  川本 千春, 矢後 亮太朗, 遠山 晏梨, 國井 理恵子, 大賀 則孝, 浅香 卓哉, 間石 奈湖, 樋田 泰浩, 樋田 京子, 北川 善政, 佐野 英彦

  The Quintessence  2020/11  クインテッセンス出版(株)
  
• The role of tumor endothelial cells in the immune environmen  [Invited]

  樋田京子

  The 79th Annual Meeting of the Japanese Cancer Assosiation Symposium, Real world of tumor microenvironment  2020/10
• 治療標的としての腫瘍血管内皮細胞の異常性  [Invited]

  樋田京子

  第79回日本癌学会学術総会特別シンポジウム「がん研究における女性研究者（WSCR）Women scientists in cancer research」  2020/10
• New Insights Into Drug-Resistance: Abnormality in Tumor Endothelial Cells  [Invited]

  樋田京子

  The 21th International Vascular Biology Meeting  2020/09
• 腫瘍血管内皮細胞の特性解析とがん治療への応用  [Invited]

  樋田京子

  第109回日本病理学会総会シンポジウム「口腔腫瘍の病理と遺伝子異常―がん形質と微小環境」  2020/04
• 腫瘍血管の多様性  [Invited]

  樋田 京子

  日本薬物動態学会第34回年会シンポジウム  2019/12
• 治療標的としての腫瘍血管の特性  [Invited]

  樋田 京子

  第60回日本脈管学会総会学術委員会シンポジウム２「がん治療～脈管を攻める、脈管を守る」  2019/10
• 歯科口腔外科学  [Not invited]

  樋田 京子

  高知大学DCセミナー  2019/10
• 血管新生とがんの悪性化  [Invited]

  樋田 京子

  第30回日本臨床口腔病理学会合同シンポジウム「口腔に症状が現れる全身疾患へのアプローチ」  2019/09
• Roles of tumor endothelial cells in tumor progression  [Invited]

  HIDA Kyoko

  the 2019 AVBS SCIENTIFIC MEETING-JOINT VASCULAR BIOLOGY MEETING  2019/09
• 腫瘍組織の多様性と相互作用  [Invited]

  樋田 京子

  文部科学省新学術領域研究 細胞社会ダイバースシテ－の統合的解明と制御_第92回日本生化学会共催シンポジウム「臓器の構築と制御の統合的理解を目指した細胞ダイバーシティー研究」  2019/09
• 腫瘍血管研究と今後の展望～Bench to Bedの実現にむけて～  [Invited]

  樋田 京子

  令和元年北海道大学歯学部口腔外科同門会学術大会  2019/09
• 間質biglycanは腫瘍血管新生を亢進し、腫瘍免疫応答を強化する(Stromal biglycan promotes tumor angiogenesis and potentiates tumor immune responses)  [Not invited]

  Li Cong, 間石 奈湖, Annan Dorcas A., 樋田 泰浩, 樋田 京子

  日本癌学会総会記事  2019/09  日本癌学会
  
• CAIIは腫瘍血管内皮細胞の増殖に重要である(Carbonic anhydrase 2(CAII) is essential for tumor endothelial cell proliferation)  [Not invited]

  間石 奈湖, Annan Dorcas A., 曽我 朋義, ダウィード・ランダ, Li Cong, 菊地 央, 北條 敬之, 森本 真弘, 北村 哲也, アラン・モハメド, 篠原 信雄, 樋田 泰浩, 樋田 京子

  日本癌学会総会記事  2019/09  日本癌学会
  
• 尿路上皮癌における抗癌剤治療後の腫瘍血管ABCB1発現亢進(Increased ABCB1 expression in tumor blood vessels of urothelial carcinoma after chemotherapy)  [Not invited]

  菊地 央, 間石 奈湖, 森本 真弘, 森本 浩史, 土屋 邦彦, 安部 崇重, 樋田 泰浩, 原林 透, 松野 吉宏, 篠原 信雄, 樋田 京子

  日本癌学会総会記事  2019/09  日本癌学会
  
• 線維芽細胞増殖因子(FGF2)は腫瘍血管内皮細胞においてTGF-βによって誘導される内皮-筋線維芽移行を制御する(Fibroblast growth factor 2 regulates TGF-β-induced endothelial-to-myofibroblast transition of tumor endothelial cells)

  吉松 康裕, 赤津 裕一, 高橋 直也, 紀室 志織, 村松 智輝, 桂 彰宏, 間石 奈湖, 鈴木 洋, 稲澤 譲治, 樋田 京子, 宮園 浩平, 渡部 徹郎

  日本癌学会総会記事  2019/09  日本癌学会
  
• Abnormalities of tumor endothelial cells and cancer progression  [Invited]

  HIDA Kyoko

  The 38th Sapporo International Cancer Symposium  2019/07
• 新たに開発した腫瘍溶解アデノウイルスの効果の検討

  加藤 泰史, 東野 史裕, 松田 彩, 間石 奈湖, 北村 哲也, 樋田 京子, 北川 善政

  日本口腔科学会雑誌  2019/07  (NPO)日本口腔科学会
  
• 腫瘍血管の多様性とその制御  [Invited]

  樋田 京子

  文部科学省科学研究費補助金 新学術領域研究「細胞社会ダイバーシティの統合的解明と制御」第四回公開シンポジウム  2019/06
• 腫瘍免疫における血管の役割  [Invited]

  樋田 京子

  第119回北海道癌談話会春季シンポジウム「がん免疫療法：基礎と臨床のコラボレーション」  2019/06
• 私のワークライフバランス－子連れ留学，ラボヘッドの経験から学んだもの  [Invited]

  樋田 京子

  第57回日本小児歯科学会大会「将来計画検討委員会・女性小児歯科医委員会合同企画」  2019/06
• VEGF阻害による腫瘍微小環境の制御  [Invited]

  樋田 京子

  Lung Cancer Expert Meeting in Sapporo,  2019/05
• 腫瘍血管内皮の遺伝子異常  [Invited]

  樋田 京子

  第108回日本病理学会総会 特別シンポジウム1「がんゲノム医療と病理」  2019/05
• 高転移性腫瘍エクソソームによる血管内皮の形質変化とがんの転移促進

  森本 真弘, 間石 奈湖, 北川 善政, 樋田 京子

  北海道歯学雑誌  2019/03  北海道歯学会
  
• 腫瘍血管新生の多様性と腎癌の悪性化  [Invited]

  樋田 京子

  腎癌の血管と免疫を見つめる会  2019/02
• 血管新生阻害と微小環境の変化  [Invited]

  HIDA Kyoko

  AIR in Sapporo－Angiogenesis & Immunology Research－  2018/11
• 腫瘍血管の特性を活かした新しいがん治療法の開発を目指して  [Invited]

  樋田 京子

  これからのキャリアを語る医師と学生の会2018講演会「『がんと遺伝子』～私たちの研究が、患者さんの未来を変える」  2018/10
• 腫瘍血管内皮細胞によるがんの転移促進  [Invited]

  HIDA Kyoko

  The 77th Annual Meeting of the Japanese Cancer Association  2018/09
• 血管新生阻害療法の展開  [Invited]

  樋田 京子

  第16回日本臨床腫瘍学会学術集会シンポジウム「消化器癌に対する血管新生阻害薬の意義」  2018/07
• 高転移性腫瘍由来エクソソームmiRNAによる血管内皮の形質変化と転移促進メカニズムの解明

  森本 真弘, 間石 奈湖, 北川 善政, 樋田 京子

  日本口腔科学会雑誌  2018/07  (NPO)日本口腔科学会
  
• 治療抵抗性における腫瘍血管の役割  [Invited]

  樋田 京子

  第107回日本病理学会総会 特別企画シンポジウム「腫瘍微小環境の病理学」  2018/06
• 化学療法誘発性IL-8による腫瘍血管におけるMDR1/ABCB1アップレギュレーション(MDR1/ABCB1 upregulation in tumor blood vessels by chemotherapy-induced IL-8)  [Not invited]

  間石 奈湖, 菊地 央, 安部 崇重, 丸山 覚, 原林 透, 飴田 要, 松野 吉宏, 樋田 泰浩, 篠原 信雄, 樋田 京子

  日本病理学会会誌  2018/04  (一社)日本病理学会
  
• 高転移性腫瘍由来エクソソームmiRNAによる血管内皮の形質変化と転移促進

  森本 真弘, 間石 奈湖, 菊地 央, 北川 善政, 樋田 京子

  日本病理学会会誌  2018/04  (一社)日本病理学会
  
• Analysis of multidrug resistant transporter expression in tumor blood vessels during chemotherapy  [Not invited]

  Maishi Nako, Kikuchi Hiroshi, Morimoto Hirofumi, Tsuchiya Kunihiko, Abe Takashige, Hida Yasuhiro, Harabayashi Toru, Matsuno Yoshihiro, Shinohara Nobuo, Hida Kyoko

  CANCER SCIENCE  2018/01
• Chemotherapy-induced drug resistance in tumor endothelial cells  [Invited]

  HIDA Kyoko

  第25回血管生物医学会学術集会 AAVBM2017シンポジウム“Tumor Angiogenesis and Lymphangiogenesis”  2017/12
• 腫瘍血管内皮細胞の形質変化とがんの悪性化  [Invited]

  樋田 京子

  第40回日本分子生物学会年会ワークショップ「生老病死における血管・リンパ管の生命科学的意義」  2017/12
• 腫瘍血管内皮細胞の特性とがんの悪性化  [Not invited]

  樋田 京子

  平成29年度先端医学研究セミナー  2017/11
• 腫瘍血管に関する基礎研究から新たながん治療戦略の構築を目指して  [Invited]

  樋田 京子

  第62回日本口腔外科学会総会・学術大会特別プログラム「日台姉妹締結10周年記念シンポジウム Translational Research」  2017/10
• 抗癌剤治療前後における腫瘍血管内皮のP-glycoprotein発現変化  [Not invited]

  間石 奈湖, 菊地 央, 森本 浩史, 土屋 邦彦, 安部 崇重, 樋田 泰浩, 原林 透, 松野 吉宏, 篠原 信雄, 樋田 京子

  日本癌学会総会記事  2017/09  日本癌学会
  
• 腫瘍血管の異常とがんの悪性化  [Invited]

  樋田 京子

  第35回日本骨代謝学会学術集会共催・第20回癌と骨病変研究会 ミニシンポジウム「癌と骨病変」  2017/07
• 腫瘍血管内皮細胞のがん幹細胞への関与  [Invited]

  樋田 京子

  第26回日本がん転移学会学術集会・総会シンポジウム2「がん幹細胞に対する新たな知見」  2017/07
• 口腔扁平上皮癌における腫瘍血管マーカーCXCR7の発現と臨床病理学的因子との比較解析

  柳谷 美沙, 間石 奈湖, 鳥居 ちさほ, 大賀 則孝, 鄭 漢忠, 進藤 正信, 北川 善政, 樋田 京子

  日本口腔科学会雑誌  2017/07  (NPO)日本口腔科学会
  
• 腫瘍血管内皮細胞の特性とがんの悪性化  [Invited]

  樋田 京子

  愛媛大学プロテオサイエンスセンター PROSセミナー&大学院特別講義  2017/06
• Analysis of multidrug resistant transporter expression in tumor blood vessels of urothelial carcinoma during chemotherapy.  [Not invited]

  Hiroshi Kikuchi, Nako Maishi, Kosuke Akiyama, Masahiro Morimoto, Misa Yanagiya, Naoto Miyajima, Kunihiko Tsuchiya, Satoshi Maruyama, Takashige Abe, Yasuhiro Hida, Toru Harabayashi, Kaname Ameda, Ryuji Matsumoto, Akira Kashiwagi, Yoshihiro Matsuno, Nobuo Shinohara, Kyoko Hida

  JOURNAL OF CLINICAL ONCOLOGY  2017/05  AMER SOC CLINICAL ONCOLOGY
  
• 抗癌剤治療前後の尿路上皮癌における腫瘍血管内皮のP-glycoprotein発現変化  [Not invited]

  菊地 央, 間石 奈湖, 土屋 邦彦, 丸山 覚, 安部 崇重, 樋田 泰浩, 原林 透, 松野 吉宏, 篠原 信雄, 樋田 京子

  日本癌学会総会記事  2016/10  日本癌学会
  
• 腫瘍血管の特異解明と新しいがん治療戦略  [Invited]

  HIDA Kyoko

  第53回大阪大学数理医学研究会  2016/09
• 腫瘍血管の特異性とエクソソーム  [Not invited]

  樋田 京子

  千里ライフサイエンスセミナーK3「エクソソーム研究の最前線：疾患のメカニズム解明から診断・治療まで」  2016/09
• 高転移性腫瘍miRによる血管内皮における薬剤耐性誘導  [Not invited]

  樋田 京子

  第25回日本がん転移学会学術集会・総会シンポジウム「がん転移の新しい治療標的としてのエクソソーム研究  2016/07
• 腫瘍血管内皮細胞の特異性解明と新たながん治療への応用  [Not invited]

  樋田 京子

  第70回日本口腔科学会学術集会「口腔疾患治療の科学革命（パラダイムシフト）  2016/04
• Heterogeneity of Tumor Endothelial Cells  [Invited]

  HIDA Kyoko

  Keystone Symposia on Molecular and Cellular Biology 2016  2016/04
• 腫瘍血管内皮細胞の特性とがんの悪性化  [Not invited]

  樋田 京子

  平成27年度「個体レベルでのがん研究支援活動」ワークショップ  2016/02
• 腫瘍血管の異常とがんの悪性化  [Invited]

  樋田 京子

  第34回日本口腔腫瘍学会総会ワークショップ  2016/01
• 腫瘍血管の遺伝子発現異常とエピゲノム  [Invited]

  樋田 京子

  第38回日本分子生物学会年会  2015/12
• 腫瘍血管の遺伝子発現異常とがんの悪性化  [Invited]

  樋田 京子

  2015/06
• 腫瘍血管の異常性獲得メカニズムについて  [Not invited]

  HIDA Kyoko

  北海道大学遺伝子病制御研究所・金沢大学がん進展制御研究所ジョイントシンポジウム2014  2014/10
• Tumor-derived exosome miRNA induce abnormality in tumor endothelial cells  [Invited]

  HIDA Kyoko

  1st JSVE annual meeting  2014/08
• 腫瘍血管の多様性と新たな治療法開発  [Invited]

  HIDA Kyoko

  金沢女性がん研究者フォーラム  2014/07
• Drug resistance in tumor blood vessels  [Not invited]

  HIDA Kyoko

  The 41st Annual Meeting of the Japanese Society of Toxicology  2014/07
• 腫瘍血管内皮の多様性解明と個別化血管新生阻害法の開発  [Invited]

  HIDA Kyoko

  公開シンポジウム「血管を標的とするナノ医療の実用化に向けて拠点形成～がんを始めとする国民病を血管から治療する～」  2014/06
• 腫瘍血管の多様性と新たな治療法開発  [Invited]

  HIDA Kyoko

  第68回NPO法人日本口腔科学会学術集会ワークショップ「がんを間質から考える」  2014/05
• Tumor endothelial cells as key players in tumor microenvironment  [Invited]

  HIDA Kyoko

  International Symposium on Vascular Biology and Medicine  2014/04
• Heterogeneity of tumor endothelium  [Invited]

  HIDA Kyoko

  The 18th International Vascular Biology Meeting  2014/04
• 腫瘍血管内皮細胞の特異性とmiRNA  [Not invited]

  HIDA Kyoko

  第36回日本分子生物学会年会ワークショップ「マイクロRNAとエクソソームの生物学的意義」  2013/12
• 腫瘍血管内皮細胞の特性解明から診断・治療への応用  [Not invited]

  HIDA Kyoko

  第3回北海道探索病理学研究シンポジウム「探索医療・基礎と臨床の連携：病理が橋渡しする臨床研究の新展開」  2013/11
• 腫瘍血管の多様性とがんの浸潤転移  [Invited]

  HIDA Kyoko

  第46回北海道病理談話会特別講演  2013/10
• 腫瘍血管の多様性  [Not invited]

  HIDA Kyoko

  第72回日本癌学会学術総会シンポジウム「がんと微小環境：研究の展望」  2013/10
• 口腔扁平上皮癌における新規予後因子Vasohibin-1の発現解析(Vasohibin-1 as a novel prognostic factor in oral squamous cell carcinoma)  [Not invited]

  鳥居 ちさほ, 大賀 則孝, 秋山 廣輔, 間石 奈湖, 北條 敬之, 大廣 洋一, 小野 貢伸, 戸塚 靖則, 鄭 漢忠, 樋田 泰浩, 進藤 正信, 佐藤 靖史, 樋田 京子

  日本癌学会総会記事  2013/10  日本癌学会
  
• 低酸素・低栄養環境によるROSの蓄積が及ぼすTECへの影響(Proangiogenic phenotype of TEC induced by ROS generation under hypoxia and starvation)  [Not invited]

  北條 敬之, 間石 奈湖, 秋山 廣輔, 大賀 則孝, 近藤 美弥子, 大村 瞳, 進藤 正信, 藤澤 俊明, 樋田 泰浩, 樋田 京子

  日本癌学会総会記事  2013/10  日本癌学会
  
• 腫瘍血管内皮におけるCXCR7の機能解析(The role of a chemokine receptor CXCR7 in tumor endothelial cells)  [Not invited]

  山田 健司, 間石 奈湖, 大賀 則孝, 秋山 廣輔, 川本 泰輔, Towfik Alam Mohammad, 進藤 正信, 高橋 典彦, 神山 俊哉, 樋田 泰浩, 武冨 紹信, 樋田 京子

  日本癌学会総会記事  2013/10  日本癌学会
  
• microvesiclesを介して腫瘍細胞由来のmiRNAが血管内皮細胞へ輸送される(Tumor-derived miRNA is transported to endothelial cells by microvesicles)  [Not invited]

  川本 泰輔, 秋山 廣輔, 大賀 則孝, 間石 奈湖, 進藤 正信, 樋田 泰浩, 小坂 展慶, 落合 孝広, 樋田 京子

  日本癌学会総会記事  2013/10  日本癌学会
  
• P-gp阻害剤はメトロノミックケモセラピーの血管新生阻害効果を強める(P-gp inhibitor enhance antiangiogenic activity in metronomic chemotherapy)  [Not invited]

  秋山 廣輔, 大賀 則孝, 樋田 泰浩, 間石 奈湖, ムハンマド・トフィック, 川本 泰輔, 大村 瞳, 山田 健司, 鳥居 ちさほ, 進藤 正信, 大場 雄介, 樋田 京子

  日本癌学会総会記事  2013/10  日本癌学会
  
• 血管内皮細胞によるがん転移促進(Endothelial cells promote tumor metastasis)  [Not invited]

  間石 奈湖, 大場 雄介, 大賀 則孝, 秋山 廣輔, 山本 和幸, 浜田 淳一, 川本 泰輔, アラム・モハメド・トウフィック, 大村 瞳, 進藤 正信, 樋田 泰浩, 樋田 京子

  日本癌学会総会記事  2013/10  日本癌学会
  
• 腫瘍血管内皮の薬剤抵抗性  [Invited]

  HIDA Kyoko

  第21回日本血管生物医学会学術総会シンポジウム  2013/09
• 腫瘍血管新生とがんの新しい治療法の開発  [Not invited]

  樋田 京子

  第25回北海道輸血シンポジウム  2013/07
• がん転移と血管新生  [Invited]

  樋田 京子

  第22回日本がん転移学会学術集会・総会  2013/07
• 泌尿器科研究は我々に何をもたらすか? 泌尿器科癌における血管新生

  大澤 崇宏, 樋田 京子, 篠原 信雄, 野々村 克也

  泌尿器外科  2013/05  医学図書出版(株)
   

  泌尿器科癌における血管新生について検討した。ヒトメラノーマ細胞(A375SM)のマウス皮下移植腫瘍からmTEC、正常皮膚からmNECを分離した。次に、ヒト腎細胞癌ならびに非癌部腎組織の血管におけるLOXの発現を解析した。A375SMのヌードマウス皮下移植モデルにおいて、LOX阻害剤BAPNを用いた治療実験を行った。LOX siRNAを用いたLOXの抑制により、mTECの遊走能、管腔形成能は有意に抑制された。また、mTECは接着斑の増加を伴いより広がった細胞へと形態変化した。さらに、LOXの抑制によりFAK tyr397のリン酸化が抑制された。BAPNを用いたヒト腫瘍のヌードマウス皮下移植モデルでは、腫瘍血管新生と肺転移が抑制された。ヒト腎癌組織と非癌部腎組織より分離培養した血管内皮において、TECにおいてLOXの高いmRNA発現レベルを認めた。
• 腫瘍血管の多様性  [Not invited]

  HIDA Kyoko

  腫瘍血管シンポジウム  2013/04
• Characterization of stem-like tumor endothelial cells, The 20th annual Meeting of the Japanese Vascular Biology and MedicineOrganization  [Not invited]

  樋田 京子

  The 10th Korea-Japan Joint Symposium on Vascular Biology  2012/12
• Heterogeneity of Tumor Endothelial Cells  [Invited]

  Kyoko HIDA

  The 43rd International Symposium of The Princess Takamatsu Cancer Research Fund“Cancer Heterogeneity: Impact on Carcinogenesis, Cancer Stem Cell, Microenvironment, Diagnosis and Treatment”  2012/11
• Altered phenotype in endothelial cells in tumor microenvironment  [Invited]

  Kyoko HIDA

  The 2nd International Symposium by JSPS Core-to-Core Program“Cooperative International Framework in TGF-β Family Signaling”  2012/10
• がん微小環境と血管新生  [Invited]

  樋田 京子

  第57回日本口腔外科学会総会・学術大会ワークショップ「がん治療戦略を目指した先端的がん研究」  2012/10
• 血管を標的とした新規抗がん剤の開発を目指して  [Invited]

  樋田 京子

  第43回PFA国際歯学会日本部会年次大会－ピエールフォーシャルアカデミー国際歯学会日本部会  2012/09
• Crosstalk between tumor endothelial cells and tumor cells  [Invited]

  Kyoko HIDA

  The 71st Annual Meeting of the Japanese Cancer Association  2012/09
• 腫瘍血管内皮細胞のプロスタサイクリン受容体は血管新生をオートクラインに促進する(Prostacyclin receptor in tumor endothelial cells promotes angiogenesis in an autocrine manner)  [Not invited]

  土屋 邦彦, 大澤 崇宏, 大賀 則孝, 樋田 泰浩, 北山 和子, 秋山 廣輔, 小野寺 雄一郎, 篠原 信雄, 藤江 学, 進藤 正信, 野々村 克也, 樋田 京子

  日本癌学会総会記事  2012/08  日本癌学会
  
• 腫瘍血管内皮におけるBiglycanの機能解析(Biglycan is a specific marker and an autocrine angiogenic factor of tumor endothelial cells)  [Not invited]

  大賀 則孝, 山本 和幸, 樋田 泰浩, 秋山 廣輔, 間石 奈湖, 川本 泰輔, 北山 和子, 大澤 崇宏, 平野 聡, 篠原 信雄, 近藤 正信, 樋田 京子

  日本癌学会総会記事  2012/08  日本癌学会
  
• がん転移における腫瘍血管内皮細胞の役割(The role of tumor endothelial cells in tumor metastasis)  [Not invited]

  間石 奈湖, 大場 雄介, 大賀 則孝, 秋山 廣輔, 山本 和幸, 浜田 淳一, 川本 泰輔, 大澤 崇宏, 近藤 美弥子, 大村 瞳, 進藤 正信, 樋田 泰浩, 樋田 京子

  日本癌学会総会記事  2012/08  日本癌学会
  
• ALDH陽性活性腫瘍血管内皮細胞の特性解析(Characterization of Aldehyde dehydrogenase (ALDH) positive tumor endothelial cells)  [Not invited]

  大村 瞳, 秋山 廣輔, 大賀 則孝, 間石 奈湖, 樋田 泰浩, 川本 泰輔, 近藤 美弥子, 大澤 崇宏, 山本 和幸, 飯田 順一, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2012/08  日本癌学会
  
• 母親として、PI（Principle Investigator）として  [Invited]

  樋田 京子

  広島大学女性研究者スキルアップ交流会  2012/07
• がんの浸潤転移と血管新生  [Invited]

  樋田 京子

  第28回日本DDS学会学術集会  2012/07
• Tumor endothelial cell s acquire drug resistance by MDR1 upregulation  [Not invited]

  Kyoko HIDA

  the 17th International Vascular Biology Meeting, 2012.6.4, Wiesbaden  2012/06
• 腫瘍血管内皮細胞の異常性と転移との関連  [Invited]

  樋田 京子

  第101回日本病理学会総会コンパニオンミーティング７「口腔癌の悪性形質の発現に関する橋渡し研究と分子標的治療への応用」  2012/04
• 血管内皮細胞の抗がん剤抵抗性  [Invited]

  樋田 京子

  千里ライフサイエンスセミナー「がんの浸潤・転移と微小環境」  2012/02
• がんの浸潤転移と血管新生  [Invited]

  樋田 京子

  第30回日本口腔腫瘍学会学術大会シンポジウム「口腔癌の浸潤－マクロ・ミクロ・モレキュラー」  2012/01
• 腫瘍血管内皮細胞の特異性の解明  [Invited]

  樋田 京子

  公開シンポジウム「血管を標的とする革新的医薬分子送達法の基盤技術の確立」  2012/01
• 腫瘍血管の多様性とがんの転移  [Invited]

  樋田 京子

  Recent Advance in Tumor Angiogenesis 2011  2011/12
• 腫瘍血管新生阻害療法における新たな局面-Lesson from Basic research  [Invited]

  樋田 京子

  函館Basic Cancer Forum  2011/11
• 腫瘍微小環境における腫瘍血管内皮異常性の獲得  [Not invited]

  樋田 京子

  第70回日本癌学会学術総会  2011/10
• 腫瘍血管内皮細胞の異常性がもたらすがん治療への影響  [Invited]

  樋田 京子

  平成23年度遺伝子病制御研究所共同研究集会、「がん細胞・組織の多様性の出現・維持に関わる微小環境因子」  2011/09
• 腫瘍血管特異性の解明と新たな血管新生阻害療法への応用  [Invited]

  樋田 京子

  大阪歯科大学大学院50周年記念講演会  2011/09
• 腫瘍血管内皮におけるLysyl oxidaseの機能解析(The role of Lysyl oxidase on abnormal phenotypes of tumor endothelial cells)  [Not invited]

  大澤 崇宏, 大賀 則孝, 北山 和子, 樋田 泰浩, 川本 泰輔, 近藤 美弥子, 間石 奈湖, 山本 和幸, 秋山 廣輔, 篠原 信雄, 野々村 克也, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2011/09  日本癌学会
  
• 転移能の異なる腫瘍から分離された腫瘍血管内皮細胞の比較解析(Comparative characterization of tumor endothelial cells isolated from highly and low metastatic tumors)  [Not invited]

  大賀 則孝, 石川 修平, 樋田 泰浩, 秋山 広輔, 間石 奈湖, 近藤 美弥子, 川本 泰輔, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2011/09  日本癌学会
  
• 腫瘍血管内皮細胞とがん転移との相互作用解析(Analysis of interaction between tumor endothelial cells and tumor metastasis)  [Not invited]

  間石 奈湖, 大賀 則孝, 樋田 泰浩, 大場 雄介, 浜田 淳一, 秋山 廣輔, 山本 和幸, 大澤 崇宏, 近藤 美弥子, 川本 泰輔, 進藤 正信, 井上 農夫男, 樋田 京子

  日本癌学会総会記事  2011/09  日本癌学会
  
• 低酸素環境が血管内皮細胞に及ぼす影響(Hypoxia is a key factor to acquisition of tumor endothelial cells abnormalities)  [Not invited]

  近藤 美弥子, 大賀 則孝, 秋山 廣輔, 間石 奈湖, 川本 泰輔, 大澤 崇宏, 山本 和幸, 大村 瞳, 樋田 泰浩, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2011/09  日本癌学会
  
• A New Insights into Tumor endothelial cell  [Invited]

  Kyoko HIDA

  The 5th Meeting of Asian Society of Oral and Maxillofacial Pathology  2011/08
• がん転移に関わる腫瘍血管内皮の異常性  [Not invited]

  樋田 京子

  第20回日本がん転移学会学術集会・総会  2011/06
• Heterogeneity of Tumor Endothelium and Drug Resistance  [Invited]

  樋田 京子

  第63回日本細胞生物学会大会、「血管の多様性～組織発生から疾患におけるダイナミクス」  2011/06
• 腫瘍血管内皮細胞の特異性とその制御  [Invited]

  樋田 京子

  ACCEL in Hokkaido～NSCLCに対するアバスチンの治療戦略～  2011/06
• 腫瘍血管内皮細胞の多様な生物像  [Invited]

  樋田 京子

  第27回日本DDS学会学術集会  2011/06
• がん組織における血管の特異性の解明と新たな血管新生阻害療法への応用  [Invited]

  樋田 京子

  高知大学医学部歯科口腔外科学講座セミナー  2011/06
• 血管の多様性 組織発生から疾患におけるダイナミクス 腫瘍血管内皮の多様性と薬剤抵抗性(Cellular and functional diversity of the vasculature: health and disease Heterogeneity of Tumor Endothelium and Drug Resistance)  [Not invited]

  樋田 京子, 大賀 則孝, 秋山 廣輔, 樋田 泰浩, 進藤 正信

  日本細胞生物学会大会講演要旨集  2011/05  (一社)日本細胞生物学会
  
• 癌間質の異常、とくに腫瘍血管の特性と浸潤との関連性  [Invited]

  樋田 京子

  第100回日本病理学会総会コンパニオンミーティング4「口腔癌の浸潤：外科病理と分子病理の架け橋」  2011/05
• がん微小環境内における腫瘍－血管の相互作用  [Invited]

  樋田 京子

  第９回口腔医科学フロンティア学術集会  2011/03
• 低酸素環境における腫瘍血管ダイナミクス  [Invited]

  樋田 京子

  The 8th Annual Meeting for the Japanese Association for Cancer and Hypoxia Research  2011/01
• Tumor induces drug resistance of endothelial cells by nuclear translocation of YB-1 and up-regulation of MDR-1  [Not invited]

  Kyoko HIDA

  第18回日本血管生物医学会学術集会  2010/12
• Crosstalk between tumor endothelial cells and microenvironment  [Invited]

  樋田 京子

  第56回日本病理学会秋期特別総会  2010/11
• Kyoko HIDA  [Invited]

  樋田 京子

  The 13th Hokkaido University – Seoul National University Joint Sympojium  2010/11
• Crosstalk between blood vessels and tumor microenvironment  [Invited]

  Kyoko HIDA

  The 13th Hokkaido University – Seoul National University Joint Sympojium  2010/10
• 腫瘍血管内皮細胞を標的とした新たな治療法の開発  [Invited]

  樋田 京子

  平成22年度北海道癌談話会秋季シンポジウム「腫瘍血管新生の基礎と臨床」  2010/10
• 癌が分泌する小胞は内皮細胞の遺伝子発現を変化させる(Tumor-derived microvesicles cause gene changes in endothelial cells)  [Not invited]

  川本 泰輔, 大賀 則孝, 北山 和子, 秋山 廣輔, 近藤 美弥子, 間石 奈湖, 大澤 崇宏, 山本 和幸, 樋田 泰浩, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2010/08  日本癌学会
  
• 転移能の異なる腫瘍から分離された腫瘍血管内皮細胞の特性解析(Comparative characterization of tumor endothelial cells isolated from highly and low metastatic tumors)  [Not invited]

  大賀 則孝, 石川 修平, 樋田 泰浩, 秋山 広輔, 北山 和子, 近藤 美弥子, 間石 奈湖, 川本 泰輔, 大澤 崇宏, 山本 和幸, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2010/08  日本癌学会
  
• がん患者末梢循環血管内皮細胞における遺伝子発現解析(Gene expression analysis of circulating endothelial cells in cancer patients)  [Not invited]

  近藤 美弥子, 大賀 則孝, 黒須 拓郎, 北山 和子, 秋山 廣輔, 間石 奈湖, 川本 泰輔, 大澤 崇宏, 山本 和幸, 樋田 泰浩, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2010/08  日本癌学会
  
• 癌細胞培養上清による正常血管内皮の薬剤抵抗性の獲得(Emdothelial cells acquire drug resistance by factors from tumor cells)  [Not invited]

  秋山 廣輔, 大賀 則孝, 樋田 泰浩, 黒須 拓郎, 北山 和子, 石川 修平, 近藤 美弥子, 間石 奈湖, 川本 泰介, 大澤 崇宏, 山本 和幸, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2010/08  日本癌学会
  
• 腫瘍血管内皮細胞と腫瘍細胞との相互作用解析(Analysis of interaction between tumor endothelial cells and tumor cells)  [Not invited]

  間石 奈湖, 大賀 則孝, 秋山 廣輔, 北山 和子, 近藤 美弥子, 川本 泰輔, 大澤 崇宏, 山本 和幸, 樋田 泰浩, 大場 雄介, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2010/08  日本癌学会
  
• 腫瘍血管内皮細胞の異常と新たな治療法の開発  [Invited]

  樋田 京子

  第64回日本口腔科学会学術集会  2010/06
• Cytogenetic Abnormalities of Tumor Endothelial Cells in Human Malignant Tumors  [Not invited]

  Kyoko HIDA

  AACR 101st ANNUAL MEETING 2010  2010/04
• 腫瘍血管内皮細胞の特性と治療抵抗性  [Invited]

  樋田 京子

  第43回加齢医学研究所シンポジウム「血管から見る医科学研究の新展開」  2010/03
• がん治療の最前線 血管新生阻害の最新アプローチ  [Invited]

  樋田 京子

  The 4th Lecture Meeting SUTENT by Urology Specialist  2010/02
• 生体ナノ環境を構成する間質細胞の時空間的ダイナミクスの解明  [Invited]

  樋田 京子

  公開シンポジウム「生体ナノ環境の時空間制御を目指して」、戦略的研究推進経費プロジェクト主催「血管を標的とする革新的医薬分子送達法の基盤技術の確立」  2010/01
• 腫瘍血管内皮細胞の異常と薬剤抵抗性  [Invited]

  樋田 京子

  第32回日本分子生物学会年会ワークショップ  2009/12
• 腫瘍血管内皮細胞の異常性  [Invited]

  樋田 京子

  第17回日本血管生物医学会  2009/10
• 腫瘍血管内皮の異常と薬剤抵抗性  [Not invited]

  樋田 京子

  第68回日本癌学会学術総会  2009/10
• 癌細胞培養上清による正常血管内皮の異常性獲得(Endothelial cells acquire abnormalities by factors from tumor cells)  [Not invited]

  秋山 廣輔, 大賀 則孝, 樋田 泰浩, 秋野 文臣, 黒須 拓郎, 石川 修平, 近藤 美弥子, 間石 奈湖, 井上 農夫男, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2009/08  日本癌学会
  
• 緑茶ポリフェノールEGCGは腫瘍血管内皮に対する抑制効果を有する(Inhibitory Effects of Epigallocatechin-3 Gallate, a Polyphenol in Green Tea, on Tumor-Associated Endothelial Cells)  [Not invited]

  大賀 則孝, 樋田 泰浩, 土屋 邦彦, 村木 力, 秋山 広輔, 近藤 美弥子, 間石 奈湖, 黒須 拓郎, 大廣 洋一, 戸塚 靖則, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2009/08  日本癌学会
  
• 転移能の異なる腫瘍から分離された腫瘍血管内皮細胞の特性解析(Comparative characterization of tumor endothelial cells isolated from high and low metastatic tumors)  [Not invited]

  石川 修平, 大賀 則孝, 樋田 泰浩, 秋野 文臣, 黒須 拓郎, 秋山 廣輔, 近藤 美弥子, 間石 奈湖, 野々村 克也, 進藤 正信, 樋田 京子

  日本癌学会総会記事  2009/08  日本癌学会
  
• がん転移におけるがん細胞・間質相互作用The Role of  [Not invited]

  樋田 京子

  第18回日本がん転移学会モーニングセッション  2009/07
• 転移能の異なる腫瘍から分離された２つの腫瘍血 管内皮細胞の比較検討  [Not invited]

  樋田 京子

  第18回日本がん転移学会  2009/07
• 腫瘍血管内皮に関する新しい洞察－がん微小環境における血管内皮の特異性  [Invited]

  樋田 京子

  北海道癌談話会春期シンポジウム  2009/06
• OP-381 腎がん腫瘍血管内皮細胞を用いた薬剤感受性の解析(腎腫瘍/基礎1,一般演題口演,第97回日本泌尿器科学会総会)

  定本 圭弘, 秋野 文臣, 樋田 京子, 篠原 信雄, 野々村 克也

  日本泌尿器科学会雑誌  2009  一般社団法人 日本泌尿器科学会
  
• がん微小環境における腫瘍血管内皮細胞の異常性  [Invited]

  樋田 京子

  日本生化学会北海道支部・日本生物物理学会北海道支部・北海道分子生物研究会２００８年度合同シンポジウム「生命現象の分子レベルでの解明」  2008/11
• 腫瘍血管内皮細胞ならびに血管内皮前駆細胞は正常血管内皮細胞に比べ、緑茶ポリフェノールに感受性を有する(Tumor endothelial cells and EPC are more sensitive to EGCG compared to normal endothelial cells)  [Not invited]

  大賀 則孝, 樋田 京子, 村木 力, 樋田 泰浩, 土屋 邦彦, 大廣 洋一, 戸塚 靖則, 進藤 正信

  日本癌学会総会記事  2008/09  日本癌学会
  
• がん微小環境における腫瘍血管内皮細胞の異常性  [Not invited]

  樋田 京子

  第73回日本インターフェロン・サイトカイン学会学術集会、第19回日本生体防御学会学術集会、第45回補体シンポジウム合同大会  2008/07
• がん微小環境における腫瘍血管内皮細胞の異常性  [Not invited]

  樋田 京子

  日本分子生物学会第８回春期シンポジウム  2008/05
• 腫瘍血管の多様性－特異的な遺伝子  [Invited]

  樋田 京子

  金沢大学がん研究所セミナー  2008/05
• 腫瘍血管新生のダイナミクス－多様な腫瘍血管内皮細胞の特性－  [Invited]

  樋田 京子

  第１回消化器リサーチセミナー  2008/04
• Chromosomal Abnormalities of Human Tumor-Associated Endothelial Cells  [Invited]

  Kyoko HIDA

  THE U.S.-JAPAN COOPERATIVE CANCER RESEARCH PROGRAM WORKSHOP 2008  2008/03
• 口腔扁平上皮がんにおけるPim-1の発現は細胞運動能を亢進し転移活性に関与する

  田中 宗一, 北村 哲也, 樋田 京子, 東野 史裕, 戸塚 靖則, 進藤 正信

  日本病理学会会誌  2008/03  (一社)日本病理学会
  
• がん微小環境における腫瘍血管内皮細胞の異常性  [Invited]

  樋田 京子

  2007日本生化・分子生物学会ワークショップ  2007/12
• Cyclooxygenase-2 inhibitorの腫瘍血管内皮ならびに血管内皮前駆細胞に対する直接的制御効果  [Not invited]

  樋田 京子

  第15回血管生物医学会学術総会  2007/11
• Abnormality of tumor endothelial cells  [Not invited]

  Kyoko HIDA

  International sessions 2 Cancer Angiogenesis.The 66th Annual Meeting of the Japanese Cancer Association  2007/10
• Abnormality of tumor endothelial cells. International sessions 2 Cancer Angiogenesis  [Invited]

  Kyoko HIDA

  The 66th Annual Meeting of the Japanese Cancer Association  2007/10
• がん微小環境における血管内皮細胞の特異性〜理想的な血管新生阻害療法を開発するために  [Invited]

  樋田 京子

  Basic Urology Research Seminar  2007/08
• 緑茶ポリフェノールによる腫瘍血管内皮と血管内皮前駆細胞への抑制効果(A green tea polyphenol, EGCG has inhibitory effects on tumor endothelial cells and endothelial progenitor cells)  [Not invited]

  大賀 則孝, 樋田 京子, 村木 力, 樋田 泰浩, 土屋 邦彦, 戸塚 靖則, 進藤 正信

  日本癌学会総会記事  2007/08  日本癌学会
  
• 腫瘍血管内皮を標的としたCyclooxigenase-2の血管新生阻害作用(Anti-Angiogenic Effects of Cyclooxygenase-2 inhibitor on Tumor-associated Endothelial Cells)  [Not invited]

  村木 力, 樋田 京子, 樋田 泰浩, 土屋 邦彦, 戸塚 靖則, Klagsbrun Michael, 進藤 正信

  日本癌学会総会記事  2007/08  日本癌学会
  
• 腫瘍血管内皮に関する新 しい洞察－がん微小環境における血管内皮の特異性  [Invited]

  樋田 京子

  北海道癌談話会春期シンポジウム  2007/06
• Abnormality of tumor-associated endothelial cells.  [Not invited]

  Kyoko HIDA

  AACR-JCA International Conference ”In the Forefront of Basic and Translational Cancer Research  2007/01
• OP-193 Adrenomedullin antagonist 発現ベクターを用いたヒト腎癌移植マウスモデルに対する遺伝子治療(第95回日本泌尿器科学会総会)

  土屋 邦彦, 樋田 京子, 村木 力, 近藤 健, 三関 哲矢, 進藤 正信, 原林 透, 篠原 信雄, 小林 正伸, 野々村 克也

  日本泌尿器科学会雑誌  2007  一般社団法人 日本泌尿器科学会
  
• 腫瘍血管内皮に関する新しい洞察  [Invited]

  樋田 京子

  東京大学大学院 医学系研究科分子病理学講座セミナー  2006/12
• Understanding Abnormality of Tumor Endothelial Cells to Develop Ideal Antiangiogenic Therapies  [Invited]

  Kyoko HIDA

  The COE 3rd International Symposium～Novel perspectives in cancer research and translation to the clinic  2006/11
• 緑茶ポリフェノールによる腫瘍血管内皮への抑制効果  [Not invited]

  樋田 京子

  第65回日本癌学会学術総会  2006/09
• Cyclooxygenase-2 inhibitorは腫瘍血管新生を抑制する  [Not invited]

  村木 力, 樋田 京子, 樋田 泰浩, 土屋 邦彦, 戸塚 靖則, Michael Klagsbrun, 進藤 正信

  日本癌学会総会記事  2006/09  日本癌学会
  
• 腫瘍血管内皮の特異的性質—染色体遺伝子学的異常について  [Not invited]

  樋田 京子

  第95回日本病理学会総会  2006/04
• 腫瘍血管内皮に関する新しい洞察－より効果的な血管新生阻害療法を目指して－  [Invited]

  樋田 京子

  旭川肺癌研究会学術大会  2006/04
• 腫瘍血管内皮についての新しい洞察  [Invited]

  樋田 京子

  第６回 愛媛オンコロジーフォーラム  2005/11
• Tumor-associated endothelial cells with cytogenetic abnormalities  [Invited]

  Kyoko HIDA

  AACR Special Conference in Cancer Research “Anti-Angiogenesis and Drug Delivery to tumors:Bench to Bedside and Back”  2005/11
• 腫瘍血管内皮についての新しい洞察  [Invited]

  樋田 京子

  第85回北海道癌談話会秋期シンポジウム「血管新生の新しいパラダイムと癌」  2005/11
• 腫瘍血管内皮の特異的性質—染色体遺伝子異常について  [Not invited]

  樋田 京子

  第50回日本口腔外科学会総会  2005/10
• 腫瘍関連血管内皮細胞は細胞発生学的に異常を伴っている  [Not invited]

  樋田 京子

  第64回日本癌学会総会  2005/09
• 腫瘍血管内皮についての新しい洞察－腫瘍血管内皮の染色体異常  [Invited]

  樋田 京子

  第85回癌談話会  2005/09
• Tumor-associated endothelial cells with cytogenetic abnormalities  [Invited]

  樋田 京子

  ボストン血管新生シンポジウム (Boston Angiogenesis Meeting: BAM)  2004/11
• Chromosome instability of Tumor associated Endothelial cells  [Invited]

  樋田 京子

  Gordon symposium  2004/08
• Tumor Endothelial cells are not normal  [Invited]

  樋田 京子

  ハーバード小児病院外科研究分野春期研究会  2003/04
• Characterization of Tumor-deirived Endothelial cells  [Invited]

  Kyoko HIda

  Harvard Medical School Surgical Research Summer Retreat Seminar  2002/07
• がん微小環境における血管の特異  [Invited]

  HIDA Kyoko

  The 104th Annual Meeting of the Japanese society of Pathology

MISC

• The functional network of biglycan: A new frontier in tumor progression

  Li Yu, Nako Maishi, Aya Matsuda, Kyoko Hida  Proteoglycan Research  1-  (3)  2023/07  

  Abstract Biglycan is a member of the small leucine‐rich proteoglycan family. Dysregulation of biglycan leads to a broad range of clinical consequences, such as osteoclastogenesis, inflammation, cardiovascular disease, and cancer. Biglycan binding to toll‐like receptor (TLR)−2 or TLR‐4 on immune cells lead to infiltration of immune cells to mediate the inflammatory response. Additionally, the extracellular matrix‐secreted soluble biglycan functions as a danger‐associated molecular pattern molecule involved in the induction of inflammation and cancer. High expression of biglycan is demonstrated in tumor endothelial cells (TECs) of various cancers and correlates with metastatic potential and poor clinical outcomes. This comprehensive review addresses the role of biglycan in both tumor cells and tumor stromal cells, especially TECs, in regulating tumor angiogenesis, tumor growth, metastasis, and chemotherapy resistance.
• Skeletal-Vascular Interactions in Bone Development, Homeostasis, and Pathological Destruction

  Haruhisa Watanabe, Nako Maishi, Marie Hoshi-Numahata, Mai Nishiura, Atsuko Nakanishi-Kimura, Kyoko Hida, Tadahiro Iimura  International Journal of Molecular Sciences  24-  (13)  10912  -10912  2023/06/30  [Refereed]
   

  Bone is a highly vascularized organ that not only plays multiple roles in supporting the body and organs but also endows the microstructure, enabling distinct cell lineages to reciprocally interact. Recent studies have uncovered relevant roles of the bone vasculature in bone patterning, morphogenesis, homeostasis, and pathological bone destruction, including osteoporosis and tumor metastasis. This review provides an overview of current topics in the interactive molecular events between endothelial cells and bone cells during bone ontogeny and discusses the future direction of this research area to find novel ways to treat bone diseases.
• Beyond starving cancer: anti-angiogenic therapy

  Kyoko Hida, Nako Maishi, Aya Matsuda, Li Yu  Journal of Medical Ultrasonics  2023/05/12  [Refereed]
  
• 洗口液中殺菌成分セチルピリジニウム塩化物水和物のSARS-CoV-2に対する効果

  武田遼, 武田遼, 間石奈湖, 松田彩, 北川善政, 樋田京子  日本口腔科学会学術集会プログラム・抄録集  77th-  2023
• Vascular inflammation induced by oral bacteria Streptococcus mutans promote tumor metastasis

  LI Y U, 間石奈湖, 赤堀永倫香, 長谷部晃, 武田遼, 松田彩, 樋田泰浩, 北川善政, 樋田京子  日本がん転移学会学術集会・総会プログラム抄録集  32nd-  2023
• Acquisition of drug resistance in endothelial cells by tumor-derived extracellular vesicles and cancer progression

  Morimoto M, Maishi N, Hida K  Cancer Drug Resistance  2023  [Refereed][Not invited]
  
• Metastatic basal cell carcinoma of buccal mucosa: a report of a rare case

  Taku Kimura, Ken-ichiro Sakata, Jun Sato, Chisato Ouchi, Noritaka Ohga, Aya Yanagawa-Matsuda, Kyoko Hida, Yoshimasa Kitagawa  World Journal of Surgical Oncology  20-  (1)  2022/12  [Refereed]
   

  Abstract Background Basal cell carcinoma (BCC) is the most common cancer worldwide. Most of BCCs can be detected in the early stages and are generally well controlled with local resection. Despite the high incidence of BCC, metastasis is rarely observed. Metastatic BCCs generally have an aggressive phenotype and are refractory to conventional treatment. Case presentation We describe a rare case of BCC in which a series of local relapses culminated in metastasis into the oral cavity 10 years after the first diagnosis of cutaneous BCC. We performed surgical resection and postoperative radiotherapy in this patient; 11 months after the final course of radiotherapy, the BCC remains stable, and the patient continues to be monitored regularly. Conclusions Because metastatic BCC is refractory to current treatment and difficult to control, his treatment history and the pathohistological features of BCC had to be considered in posttreatment planning.
• 口底に認めた脂肪腫の1例

  中村 圭佑, 坂田 健一郎, 佐藤 淳, 羽藤 裕之, 村井 知佳, 大賀 則孝, 松田 彩, 間石 奈湖, 樋田 京子, 北川 善政  北海道歯学雑誌  43-  88  -93  2022/09  

  【緒言】口腔領域における脂肪腫の発生頻度は全体の約2%と報告されている.発生部位は頬粘膜,舌,口唇,歯肉の順に多く口底での発生は比較的まれで,2cm以下が半数を占める.著者らは認知症を伴う70歳代女性の口底脂肪腫を経験した.【症例と経過】近医歯科より口底部腫瘤の精査を勧められ,当科を紹介された.左側口底部に30×25mm大の腫瘤病変を認めた.表面粘膜は滑沢で,境界明瞭,弾性軟で,粘膜下の腫瘤の一部は黄色を呈していた.病変はCTで境界明瞭な低吸収域,USでは高エコー領域を認めた.認知症および腫瘤病変により口腔清掃不良状態で,残根の歯を多数認めた.口底良性腫瘍の臨床診断の下,腫瘍摘出術を施行した.摘出物は充実性で,薄い披膜に包まれていた.病理組織学的診断は脂肪腫であった.【結語】本病変の増大時期は認知症のため不明であったが,腫瘤の増大により口腔清掃不良を引き起こしていた.症例によっても異なるが脂肪腫を含め口腔内の環境を悪化させている原因は可能な限り早期の手術が望ましいと考えられた.(著者抄録)
• The Roles of Tumor Endothelial Cells in Cancer Metastasis

  Hida K, Maishi N, Takeda R, Hida Y  Metastasis  2022/03  [Not refereed][Not invited]
  
• SARS-CoV-2感染マウスを用いた肺血管内皮細胞の重症化関連遺伝子の解析

  武田遼, 武田遼, 伊藤航, 手代木孝仁, 佐々木道仁, 大場靖子, 大場靖子, 間石奈湖, 松田彩, 佐藤彰彦, 鳥羽晋輔, 李智媛, 飯村忠浩, 樋田泰浩, 北川善政, 澤洋文, 澤洋文, 澤洋文, 樋田京子  日本血管生物医学会学術集会プログラム・抄録集  30th (CD-ROM)-  2022
• Association between high endothelial venules and clinical factors in oral squamous cell carcinoma

  Takashi Niiyama, Nako Maishi, Aya Matsuda, Kyoko Hida  CANCER SCIENCE  112-  865  -865  2021/02
• 腫瘍血管内皮細胞における異常性解明とその制御

  間石奈湖, 樋田京子  高倉伸幸編「細胞」－特集 がんの進展における血管新生と免疫－  53-  (2)  49  -52  2021/02  [Not refereed][Not invited]
  
• 腫瘍血管新生因子Biglycanを標的とした阻害剤探索系の構築

  石井豪, 間石奈湖, 積田卓也, 竹川英輝, 梅山悠伊, 樋田京子, 竹田浩之  日本生化学会大会(Web)  94th-  2021
• A case of extensive exostoses in the maxilla and mandible managed with model surgery of the postoperative denture spaces

  馬場陽久, 岡田和隆, 坂田健一郎, 羽藤裕之, 松田彩, 樋田京子, 北川善政, 山崎裕  北海道歯学雑誌  42-  2021
• がん治療標的としての腫瘍血管内皮細胞の特異性

  間石奈湖, Annan DA, 樋田京子  炎症と免疫  28-  (5)  35  -40  2020/08  [Not refereed][Not invited]
  
• Trial to reinforce the effect of our oncolytic adenovirus

  松田彩, 加藤泰史, 北村哲也, 間石奈湖, 樋田京子, 東野史裕  日本病理学会会誌  109-  (1)  2020
• 線維芽細胞増殖因子(FGF2)は腫瘍血管内皮細胞においてTGF-βによって誘導される内皮-筋線維芽移行を制御する(Fibroblast growth factor 2 regulates TGF-β-induced endothelial-to-myofibroblast transition of tumor endothelial cells)

  吉松 康裕, 赤津 裕一, 高橋 直也, 紀室 志織, 村松 智輝, 桂 彰宏, 間石 奈湖, 鈴木 洋, 稲澤 譲治, 樋田 京子, 宮園 浩平, 渡部 徹郎  日本癌学会総会記事  78回-  P  -2280  2019/09
• 尿路上皮癌における抗癌剤治療後の腫瘍血管ABCB1発現亢進(Increased ABCB1 expression in tumor blood vessels of urothelial carcinoma after chemotherapy)

  菊地 央, 間石 奈湖, 森本 真弘, 森本 浩史, 土屋 邦彦, 安部 崇重, 樋田 泰浩, 原林 透, 松野 吉宏, 篠原 信雄, 樋田 京子  日本癌学会総会記事  78回-  P  -1267  2019/09  [Refereed][Not invited]
  
• CAIIは腫瘍血管内皮細胞の増殖に重要である(Carbonic anhydrase 2(CAII) is essential for tumor endothelial cell proliferation)

  間石 奈湖, Annan Dorcas A., 曽我 朋義, ダウィード・ランダ, Li Cong, 菊地 央, 北條 敬之, 森本 真弘, 北村 哲也, アラン・モハメド, 篠原 信雄, 樋田 泰浩, 樋田 京子  日本癌学会総会記事  78回-  E  -2101  2019/09  [Refereed][Not invited]
  
• 間質biglycanは腫瘍血管新生を亢進し、腫瘍免疫応答を強化する(Stromal biglycan promotes tumor angiogenesis and potentiates tumor immune responses)

  Li Cong, 間石 奈湖, Annan Dorcas A., 樋田 泰浩, 樋田 京子  日本癌学会総会記事  78回-  P  -2247  2019/09  [Refereed][Not invited]
  
• 破骨細胞の分化・機能とRNA結合タンパクHuRの細胞質局在

  養田稔, 養田稔, 伊藤啓介, 松田彩, 間石奈湖, 北村哲也, 樋田京子, 鄭漢忠, 東野史裕  日本病理学会会誌  108-  (1)  2019
• 尿路上皮癌における抗癌剤治療後の腫瘍血管ABCB1発現亢進と薬剤耐性

  菊地 央, 間石 奈湖, 宮田 遥, 松本 隆児, 大澤 孝宏, 安部 崇重, 樋田 泰浩, 丸山 覚, 原林 透, 飴田 要, 柏木 明, 松野 吉宏, 篠原 信雄, 樋田 京子  日本癌治療学会学術集会抄録集  56回-  O22  -3  2018/10  [Not refereed][Not invited]
  
• 化学療法誘発性IL-8による腫瘍血管におけるMDR1/ABCB1アップレギュレーション(MDR1/ABCB1 upregulation in tumor blood vessels by chemotherapy-induced IL-8)

  間石 奈湖, 菊地 央, 安部 崇重, 丸山 覚, 原林 透, 飴田 要, 松野 吉宏, 樋田 泰浩, 篠原 信雄, 樋田 京子  日本病理学会会誌  107-  (1)  321  -321  2018/04  [Refereed][Not invited]
  
• 肺癌症例における腫瘍血管内皮マーカーbiglycanの発現解析

  森本 浩史, 間石 奈湖, 樋田 泰浩, 西原 広史, 畑中 豊, 松野 吉宏, 中村 透, 平野 聡, 樋田 京子  日本病理学会会誌  107-  (1)  293  -293  2018/04  [Not refereed][Not invited]
  
• 肺癌症例における腫瘍血管内皮マーカーbiglycanの発現解析

  森本 浩史, 間石 奈湖, 樋田 泰浩, 西原 広史, 畑中 豊, 松野 吉宏, 中村 透, 平野 聡, 樋田 京子  日本病理学会会誌  107-  (1)  293  -293  2018/04  [Not refereed][Not invited]
  
• Analysis of multidrug resistant transporter expression in tumor blood vessels during chemotherapy

  Maishi Nako, Kikuchi Hiroshi, Morimoto Hirofumi, Tsuchiya Kunihiko, Abe Takashige, Hida Yasuhiro, Harabayashi Toru, Matsuno Yoshihiro, Shinohara Nobuo, Hida Kyoko  CANCER SCIENCE  109-  873  2018/01  [Refereed][Not invited]
  
• 腫瘍血管内皮細胞の特異性

  HIDA Kyoko  Journal of the Society of Japanese Women Scientists  18-  12  -17  2018  [Refereed][Not invited]
  
• 抗癌剤治療前後における腫瘍血管内皮のP-glycoprotein発現変化

  間石 奈湖, 菊地 央, 森本 浩史, 土屋 邦彦, 安部 崇重, 樋田 泰浩, 原林 透, 松野 吉宏, 篠原 信雄, 樋田 京子  日本癌学会総会記事  76回-  J  -3103  2017/09  [Refereed][Not invited]
  
• 肺癌症例における腫瘍血管内皮細胞マーカーbiglycanの発現解析

  森本 浩史, 間石 奈湖, 樋田 泰浩, 西原 広史, 畑中 豊, 松野 吉宏, 中村 透, 平野 聡, 樋田 京子  日本癌学会総会記事  76回-  P  -2436  2017/09  [Not refereed][Not invited]
  
• 肺癌症例における腫瘍血管内皮細胞マーカーbiglycanの発現解析

  森本 浩史, 間石 奈湖, 樋田 泰浩, 西原 広史, 畑中 豊, 松野 吉宏, 中村 透, 平野 聡, 樋田 京子  日本癌学会総会記事  76回-  P  -2436  2017/09  [Not refereed][Not invited]
  
• 抗癌剤治療前後における腫瘍血管内皮のP-glycoprotein発現変化

  間石 奈湖, 菊地 央, 森本 浩史, 土屋 邦彦, 安部 崇重, 樋田 泰浩, 原林 透, 松野 吉宏, 篠原 信雄, 樋田 京子  日本癌学会総会記事  76回-  J  -3103  2017/09  [Not refereed][Not invited]
  
• 【血管制御系と疾患】 腫瘍微小環境における血管系の異常

  樋田 京子, 間石 奈湖  生体の科学  68-  (4)  321  -323  2017/08  [Not refereed][Not invited]
  
• 肺癌症例における腫瘍血管内皮マーカーbiglycanの発現解析

  森本浩史, 間石奈湖, 樋田泰浩, 西原広史, 畑中豊, 松野吉宏, 平野聡, 樋田京子  がんと代謝研究会プログラム&抄録集  5th-  74  2017/07  [Not refereed][Not invited]
  
• Analysis of multidrug resistant transporter expression in tumor blood vessels of urothelial carcinoma during chemotherapy.

  Hiroshi Kikuchi, Nako Maishi, Kosuke Akiyama, Masahiro Morimoto, Misa Yanagiya, Naoto Miyajima, Kunihiko Tsuchiya, Satoshi Maruyama, Takashige Abe, Yasuhiro Hida, Toru Harabayashi, Kaname Ameda, Ryuji Matsumoto, Akira Kashiwagi, Yoshihiro Matsuno, Nobuo Shinohara, Kyoko Hida  JOURNAL OF CLINICAL ONCOLOGY  35-  2017/05  [Refereed][Not invited]
  
• 尿路上皮癌における抗癌剤治療後の腫瘍血管ABCB1発現亢進

  菊地央, 菊地央, 間石奈湖, 秋山廣輔, 森本真弘, 土屋邦彦, 丸山覚, 安部崇重, 樋田泰浩, 原林透, 飴田要, 松本隆児, 松本隆児, 柏木明, 松野吉宏, 篠原信雄, 樋田京子  日本がん転移学会学術集会・総会プログラム抄録集  26th-  134  2017  [Not refereed][Not invited]
  
• 抗癌剤治療前後の尿路上皮癌における腫瘍血管内皮のP-glycoprotein発現変化

  菊地 央, 間石 奈湖, 土屋 邦彦, 丸山 覚, 安部 崇重, 樋田 泰浩, 原林 透, 松野 吉宏, 篠原 信雄, 樋田 京子  日本癌学会総会記事  75回-  P  -3248  2016/10  [Refereed][Not invited]
  
• 腫瘍血管・腫瘍血管内皮細胞の特徴とその分子機構

  間石奈湖, 樋田京子  医学のあゆみ  258-  (1)  11  -14  2016/07  [Not refereed][Not invited]
  
• マイクロRNAと血管疾患

  樋田 京子  血管医学：今が旬！マイクロRNA・エクソソーム最前線  17-  (1)  71  -76  2016/03  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞の多様性

  HIDA KYOKO, OGA NORITAKA, OGA NORITAKA, MAISHI NAKO, AKIYAMA KOSUKE, HIDA YASUHIRO  Bio Clin  5-  (1)  158  -164  2016/02  [Not refereed][Not invited]
   

  腫瘍血管内皮細胞は正常血管内皮細胞と共通の性質をもつと近年まで考えられてきたが、腫瘍血管は形態学的異常のみならず、内皮細胞にも異常性があることがわかってきた。一方、がん微小環境には多様性があることが明らかになってきた。本研究では転移能の異なる2種類の腫瘍に着目し、それらの腫瘍から分離培養された腫瘍血管内皮細胞には様々な違いがあることを示した。このことにより、微小環境が腫瘍血管内皮細胞の特異性獲得の原因になっていることが示唆された。(著者抄録)
• 腫瘍血管の分子生物学的特徴

  樋田 京子  細胞工学  35-  (1)  19  -22  2016/01  [Not refereed][Not invited]
  
• 高転移性腫瘍miRNAはIL-6ならびにALDHの発現亢進を介して血管内皮に薬剤耐性を誘導する

  鳥居 ちさほ, 秋山 廣輔, 川本 泰輔, 間石 奈湖, 鄭 漢忠, 戸塚 靖則, 進藤 正信, 樋田 泰浩, 南 敬, 小坂 展慶, 落谷 孝広, 樋田 京子  日本癌学会総会記事  74回-  P  -3069  2015/10  [Refereed][Not invited]
  
• ROSによるbiglycan発現誘導を介した腫瘍血管内皮細胞の血管新生能亢進

  北條 敬之, 間石 奈湖, Alam Mohammad T., 秋山 廣輔, 大賀 則孝, 進藤 正信, 樋田 泰浩, 藤澤 俊明, 樋田 京子  日本癌学会総会記事  74回-  P  -3106  2015/10  [Refereed][Not invited]
  
• 高転移性腫瘍miRNAはIL-6ならびにALDHの発現亢進を介して血管内皮に薬剤耐性を誘導する

  鳥居 ちさほ, 秋山 廣輔, 川本 泰輔, 間石 奈湖, 鄭 漢忠, 戸塚 靖則, 進藤 正信, 樋田 泰浩, 南 敬, 小坂 展慶, 落谷 孝広, 樋田 京子  日本癌学会総会記事  74回-  P  -3069  2015/10  [Not refereed][Not invited]
  
• CRKアダプター蛋白質はHGF/c-Metフィードバックループを介して膀胱癌のEMTと転移を誘導する

  王 磊, 松本 隆児, 津田 真寿美, 間石 奈湖, 安部 崇重, 木村 太一, 谷野 美智枝, 西原 広史, 樋田 京子, 大場 雄介, 篠原 信雄, 田中 伸哉  日本癌学会総会記事  74回-  J  -1142  2015/10  [Not refereed][Not invited]
  
• 腫瘍微小環境において、腫瘍由来のmiRXにより血管内皮はIL-6をオートクラインで利用し、薬剤耐性や幹細胞性を獲得している

  鳥居 ちさほ, 秋山 廣輔, 川本 泰輔, 間石 奈湖, 鄭 漢忠, 進藤 正信, 樋田 泰浩, 樋田 京子  日本口腔科学会雑誌  64-  (2)  191  -191  2015/07  [Refereed][Not invited]
  
• がんにおける血管・リンパ管新生 5.腫瘍血管の特性を標的とした新たな治療法の展望

  HIDA KYOKO, MAISHI NAKO, HIDA YASUHIRO  実験医学  33-  (5)  794  -798  2015/03/15  [Not refereed][Not invited]
  
• 腫瘍血管の特性を標的とした新たな治療法の展望

  樋田京子, 間石奈湖, 樋田泰浩  実験医学 増刊「がん微小環境と標的治療」  33-  (5)  134  -138  2015  [Not refereed][Not invited]
  
• CXCR7を標的とした新規血管新生阻害療法の可能性(CXCR7 as a novel target for anti-angiogenic therapy)

  山田 健司, 間石 奈湖, 秋山 廣輔, 大賀 則孝, 川本 泰輔, 進藤 正信, 高橋 典彦, 神山 俊哉, 樋田 泰浩, 武冨 紹信, 樋田 京子  日本癌学会総会記事  73回-  J  -1006  2014/09  [Refereed][Not invited]
  
• 腫瘍内低酸素環境と腫瘍血管内皮の異常性の関連

  大賀 則孝, 近藤 美弥子, 秋山 廣輔, 間石 奈湖, 北川 善政, 進藤 正信, 樋田 京子  日本口腔科学会雑誌  63-  (4)  347  -347  2014/09  [Not refereed][Not invited]
  
• OP-027-5 腫瘍血管内皮におけるCXCR7の機能解析(OP-027 基礎 腫瘍免疫・発癌・その他,一般演題,第114回日本外科学会定期学術集会)

  山田 健司, 間石 奈湖, 大賀 則孝, 秋山 廣輔, 樋田 泰浩, 川本 泰輔, Towfik Alam Mohammad, 高橋 典彦, 神山 俊哉, 樋田 京子, 武冨 紹信  日本外科学会雑誌  115-  (2)  361  -361  2014/03/05
• 腫瘍血管内皮におけるCXCR7の機能解析

  山田健司, 間石奈湖, 大賀則孝, 秋山廣輔, 樋田泰浩, 川本泰輔, MOHAMMAD Towfik Alam, 高橋典彦, 神山俊哉, 樋田京子, 武冨紹信  日本外科学会雑誌  115-  361  2014/03/05  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞のがん転移促進

  間石奈湖, 大場雄介, 大賀則孝, 大賀則孝, 秋山廣輔, 浜田淳一, 山本和幸, 川本泰輔, 進藤正信, 樋田泰浩, 樋田京子  日本病理学会会誌  103-  (1)  209  -209  2014/03  [Not refereed][Not invited]
  
• 口腔扁平上皮癌における新規予後因子としてのVasohibin‐1

  鳥居ちさほ, 鳥居ちさほ, 大賀則孝, 大賀則孝, 秋山廣輔, 間石奈湖, 北條敬之, 大廣洋一, 小野貢伸, 戸塚靖則, 北川善正, 鄭漢忠, 樋田泰浩, 進藤正信, 松野吉宏, 佐藤靖史, 樋田京子  日本病理学会会誌  103-  (1)  281  -281  2014/03  [Not refereed][Not invited]
  
• 腫瘍血管における新規血管新生関連因子Biglycanの発現と機能

  安永賢史, 間石奈湖, 秋山廣輔, 山本和幸, MOHAMMAD Alam T, 大賀則孝, 大賀則孝, 篠原信雄, 進藤正信, 樋田泰浩, 樋田京子  日本病理学会会誌  103-  (1)  397  -397  2014/03  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞のがん転移への関わり

  間石奈湖, 大場雄介, 樋田泰浩, 浜田淳一, 山本和幸, 大賀則孝, 秋山廣輔, 川本泰輔, 大澤崇宏, 近藤美弥子, 大村瞳, 井上農夫男, 進藤正信, 樋田京子  日本口腔科学会雑誌  63-  (1)  135  -135  2014/01  [Not refereed][Not invited]
  
• 転移能の異なる腫瘍由来の血管内皮細胞の特性解析

  間石奈湖, 大賀則孝, 石川修平, 樋田泰浩, 秋山廣輔, 川本泰輔, 進藤正信, 樋田京子  日本口腔科学会雑誌  63-  (1)  92  -92  2014/01  [Not refereed][Not invited]
  
• ALDH陽性腫瘍血管内皮細胞の特性解析

  大村瞳, 秋山廣輔, 大賀則孝, 間石奈湖, TOWFIK Alam Mohammad, 樋田泰浩, 川本泰輔, 近藤美弥子, 大澤崇宏, 飯田順一郎, 進藤正信, 樋田京子  日本口腔科学会雑誌  63-  (1)  133  -133  2014/01  [Not refereed][Not invited]
  
• ベラパミルはパクリタキセルの抗腫瘍効果を増強する

  秋山廣輔, 大賀則孝, 樋田泰浩, 間石奈湖, TOWFIK Alam Mohammad, 川本泰輔, 大村瞳, 山田健司, 鳥居ちさほ, 進藤正信, 樋田京子  日本口腔科学会雑誌  63-  (1)  123  -123  2014/01  [Not refereed][Not invited]
  
• 腫瘍細胞由来のmicrovesicleが血管内皮細胞のproangiogenic phenotypeを引き起こす

  秋山廣輔, 川本泰輔, 大賀則孝, 樋田泰浩, 近藤美弥子, 間石奈湖, 大澤崇宏, 山本和幸, 進藤正信, 樋田京子  日本口腔科学会雑誌  63-  (1)  170  -170  2014/01  [Not refereed][Not invited]
  
• 腫瘍血管新生阻害療法の今後の展望

  樋田 京子  Surgery Frontier～特集：癌と微小環境  21-  (2)  53  -57  2014  [Not refereed][Not invited]
  
• 腫瘍血管内皮の特異性と新たな治療戦略への展望

  HIDA Kyoko  病理と臨床～特集：がんと間質  32-  (1)  45  -50  2014  [Not refereed][Not invited]
  
• Therapeutic Sensitivity of Tumor Endothelial Cells

  秋山 廣輔, 樋田 京子  最新医学  68-  (12)  2626  -2631  2013/12
• 腫瘍血管内皮の特異性と新規治療への応用

  HIDA Kyoko  がん分子標的治療 別冊  11-  (2)  55  -58  2013/12  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞の特異性と治療感受性

  HIDA Kyoko  最新医学  68-  (12)  40  -45  2013/12  [Not refereed][Not invited]
  
• 血管内皮細胞によるがん転移促進(Endothelial cells promote tumor metastasis)

  間石 奈湖, 大場 雄介, 大賀 則孝, 秋山 廣輔, 山本 和幸, 浜田 淳一, 川本 泰輔, アラム・モハメド・トウフィック, 大村 瞳, 進藤 正信, 樋田 泰浩, 樋田 京子  日本癌学会総会記事  72回-  127  -127  2013/10  [Refereed][Not invited]
  
• P-gp阻害剤はメトロノミックケモセラピーの血管新生阻害効果を強める(P-gp inhibitor enhance antiangiogenic activity in metronomic chemotherapy)

  秋山 廣輔, 大賀 則孝, 樋田 泰浩, 間石 奈湖, ムハンマド・トフィック, 川本 泰輔, 大村 瞳, 山田 健司, 鳥居 ちさほ, 進藤 正信, 大場 雄介, 樋田 京子  日本癌学会総会記事  72回-  127  -127  2013/10  [Refereed][Not invited]
  
• microvesiclesを介して腫瘍細胞由来のmiRNAが血管内皮細胞へ輸送される(Tumor-derived miRNA is transported to endothelial cells by microvesicles)

  川本 泰輔, 秋山 廣輔, 大賀 則孝, 間石 奈湖, 進藤 正信, 樋田 泰浩, 小坂 展慶, 落合 孝広, 樋田 京子  日本癌学会総会記事  72回-  267  -267  2013/10  [Refereed][Not invited]
  
• 腫瘍血管内皮におけるCXCR7の機能解析(The role of a chemokine receptor CXCR7 in tumor endothelial cells)

  山田 健司, 間石 奈湖, 大賀 則孝, 秋山 廣輔, 川本 泰輔, Towfik Alam Mohammad, 進藤 正信, 高橋 典彦, 神山 俊哉, 樋田 泰浩, 武冨 紹信, 樋田 京子  日本癌学会総会記事  72回-  317  -317  2013/10  [Refereed][Not invited]
  
• 低酸素・低栄養環境によるROSの蓄積が及ぼすTECへの影響(Proangiogenic phenotype of TEC induced by ROS generation under hypoxia and starvation)

  北條 敬之, 間石 奈湖, 秋山 廣輔, 大賀 則孝, 近藤 美弥子, 大村 瞳, 進藤 正信, 藤澤 俊明, 樋田 泰浩, 樋田 京子  日本癌学会総会記事  72回-  317  -317  2013/10  [Refereed][Not invited]
  
• 口腔扁平上皮癌における新規予後因子Vasohibin-1の発現解析(Vasohibin-1 as a novel prognostic factor in oral squamous cell carcinoma)

  鳥居 ちさほ, 大賀 則孝, 秋山 廣輔, 間石 奈湖, 北條 敬之, 大廣 洋一, 小野 貢伸, 戸塚 靖則, 鄭 漢忠, 樋田 泰浩, 進藤 正信, 佐藤 靖史, 樋田 京子  日本癌学会総会記事  72回-  414  -414  2013/10  [Refereed][Not invited]
  
• 腫瘍血管内皮細胞のがん転移への関与

  間石奈湖, 大場雄介, 山本和幸, 大賀則孝, 浜田淳一, 秋山廣輔, 川本泰輔, 大澤崇宏, 大村瞳, 樋田泰浩, 進藤正信, 樋田京子  日本病理学会会誌  102-  (1)  437  2013/04/26  [Not refereed][Not invited]
  
• 腫瘍血管内皮におけるCXCR7の機能解析

  山田健司, 間石奈湖, 大賀則考, 秋山廣輔, 川本泰輔, TOWFIK Mohammad, 大村瞳, 鳥居ちさほ, 高橋典彦, 樋田泰浩, 進藤正信, 武冨紹信, 樋田京子  日本病理学会会誌  102-  (1)  307  2013/04/26  [Not refereed][Not invited]
  
• Biglycan は腫瘍血管内皮における新規血管新生関連因子である

  山本 和幸, 大賀 則孝, 樋田 泰浩, 間石 奈湖, 川本 泰輔, 秋山 廣輔, 大澤 崇宏, 近藤 美弥子, 加賀 基知三, 平野 聡, 篠原 信雄, 進藤 正信, 樋田 京子  北海道醫學雜誌 = Acta medica Hokkaidonensia  88-  (2)  103  -103  2013/04/01  [Not refereed][Not invited]
  
• Biglycanは腫瘍血管における新規血管新生関連因子である

  TOWFIK Alam, 大賀則孝, 山本和幸, 秋山廣輔, 間石奈湖, 川本泰輔, 進藤正信, 樋田泰浩, 樋田京子  日本病理学会会誌  102-  (1)  437  -437  2013/04  [Not refereed][Not invited]
  
• 腫瘍血管を標的としたMetronomic ChemotherapyとP‐gp阻害剤の併用療法の有効性の検討

  秋山廣輔, 大賀則孝, 樋田泰浩, 間石奈湖, TOWFIK Mohammad Alam, 川本泰輔, 大村瞳, 山田健司, 鳥居ちさほ, 進藤正信, 樋田京子  日本病理学会会誌  102-  (1)  325  -325  2013/04  [Not refereed][Not invited]
  
• ALDH陽性腫瘍血管内皮細胞の特性解析

  大村瞳, 秋山廣輔, 大賀則孝, 間石奈湖, TOWFIK Alam Mohammad, 樋田泰浩, 川本泰輔, 近藤美弥子, 大澤崇宏, 飯田順一郎, 進藤正信, 樋田京子  日本病理学会会誌  102-  (1)  435  -435  2013/04  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞の骨分化能

  樋田泰浩, 大賀則孝, 秋山廣輔, 間石奈湖, 川本泰輔, 大村瞳, 鳥居さちほ, 松居喜郎, 樋田京子  再生医療  12-  249  2013/02/28  [Not refereed][Not invited]
  
• P‐gp阻害剤はパクリタキセルの抗腫瘍効果を増強する

  秋山廣輔, 大賀則孝, 樋田泰浩, 間石奈湖, TOWFIK Alam Mohammad, 川本泰輔, 大村瞳, 山田健司, 鳥居ちさほ, 進藤正信, 樋田京子  日本がん転移学会学術集会・総会プログラム抄録集  22nd-  78  2013  [Not refereed][Not invited]
  
• がん転移における腫瘍血管内皮細胞の役割

  間石奈湖, 大場雄介, 大賀則孝, 秋山廣輔, 山本和幸, 浜田淳一, 川本泰輔, ALAM Mohammad T, 樋田泰浩, 樋田京子  日本血管生物医学会学術集会プログラム・抄録集  21st-  134  2013  [Not refereed][Not invited]
  
• がん微小環境における腫瘍血管の異常性獲得機構の解明

  樋田京子, 大賀則孝, 秋山廣輔, 間石奈湖, 川本泰輔, 大場雄介, 樋田泰浩  がん研究分野の特性等を踏まえた支援活動公開シンポジウムプログラム・抄録集 平成24年度  32  2013  [Not refereed][Not invited]
  
• 低酸素・低栄養により誘導されるROSが及ぼす腫瘍血管内皮への影響

  北條敬之, 間石奈湖, 秋山廣輔, 大賀則孝, 近藤美弥子, 角谷瞳, 進藤正信, 樋田泰浩, 藤澤俊明, 樋田京子  日本血管生物医学会学術集会プログラム・抄録集  21st-  154  2013  [Not refereed][Not invited]
  
• がん転移における腫瘍血管内皮細胞の役割

  間石奈湖, 大場雄介, 大賀則孝, 秋山廣輔, 山本和幸, 浜田淳一, 川本泰輔, ALAM MOHAMMAD T, 大村瞳, 樋田泰浩, 樋田京子  日本がん転移学会学術集会・総会プログラム抄録集  22nd-  79  2013  [Not refereed][Not invited]
  
• 腫瘍微小環境におけるVEGFシグナルを介したMDR1の発現亢進による血管内皮細胞の薬剤抵抗性獲得について

  秋山廣輔, 大賀則孝, 樋田泰浩, 川本泰輔, 定本圭弘, 石川修平, 間石奈湖, 間石奈湖, 秋野文臣, 近藤美弥子, 近藤美弥子, 松田彩, 井上農夫男, 進藤正信, 樋田京子  日本口腔科学会雑誌  62-  (1)  169  -169  2013/01  [Not refereed][Not invited]
  
• 腫瘍血管内皮におけるCXCR7の機能解析

  山田健司, 間石奈湖, 大賀則孝, 秋山廣輔, 樋田泰浩, 川本泰輔, ALAM Mohammad Towfik, 高橋典彦, 神山俊哉, 武冨紹信, 樋田京子  日本血管生物医学会学術集会プログラム・抄録集  21st-  155  2013  [Not refereed][Not invited]
  
• ALDH陽性活性腫瘍血管内皮細胞の特性解析(Characterization of Aldehyde dehydrogenase (ALDH) positive tumor endothelial cells)

  大村 瞳, 秋山 廣輔, 大賀 則孝, 間石 奈湖, 樋田 泰浩, 川本 泰輔, 近藤 美弥子, 大澤 崇宏, 山本 和幸, 飯田 順一, 進藤 正信, 樋田 京子  日本癌学会総会記事  71回-  85  -85  2012/08  [Refereed][Not invited]
  
• がん転移における腫瘍血管内皮細胞の役割(The role of tumor endothelial cells in tumor metastasis)

  間石 奈湖, 大場 雄介, 大賀 則孝, 秋山 廣輔, 山本 和幸, 浜田 淳一, 川本 泰輔, 大澤 崇宏, 近藤 美弥子, 大村 瞳, 進藤 正信, 樋田 泰浩, 樋田 京子  日本癌学会総会記事  71回-  85  -85  2012/08  [Refereed][Not invited]
  
• 腫瘍血管内皮におけるBiglycanの機能解析(Biglycan is a specific marker and an autocrine angiogenic factor of tumor endothelial cells)

  大賀 則孝, 山本 和幸, 樋田 泰浩, 秋山 廣輔, 間石 奈湖, 川本 泰輔, 北山 和子, 大澤 崇宏, 平野 聡, 篠原 信雄, 近藤 正信, 樋田 京子  日本癌学会総会記事  71回-  86  -86  2012/08  [Refereed][Not invited]
  
• 腫瘍血管内皮細胞のプロスタサイクリン受容体は血管新生をオートクラインに促進する(Prostacyclin receptor in tumor endothelial cells promotes angiogenesis in an autocrine manner)

  土屋 邦彦, 大澤 崇宏, 大賀 則孝, 樋田 泰浩, 北山 和子, 秋山 廣輔, 小野寺 雄一郎, 篠原 信雄, 藤江 学, 進藤 正信, 野々村 克也, 樋田 京子  日本癌学会総会記事  71回-  251  -251  2012/08  [Refereed][Not invited]
  
• 腫瘍細胞由来のmicrovesiclesがエンドサイトーシスを介して血管内皮細胞に血管新生能を獲得させる(Tumor-derived microvesicles induce proangiogenic phenotype in endothelial cells via endocytosis)

  川本 泰輔, 大賀 則孝, 秋山 廣輔, 平田 尚也, 北原 秀治, 間石 奈湖, 大澤 崇宏, 山本 和幸, 近藤 美弥子, 進藤 正信, 樋田 泰浩, 樋田 京子  日本癌学会総会記事  71回-  293  -293  2012/08  [Not refereed][Not invited]
  
• 転移能の異なる腫瘍由来の血管内皮細胞の特性解析

  大賀則孝, 石川修平, 樋田泰浩, 秋山廣輔, 間石奈湖, 川本泰輔, 進藤正信, 樋田京子  日本DDS学会学術集会プログラム予稿集  28回-  173  -173  2012/06  [Not refereed][Not invited]
  
• Biglyanは,腫瘍血管内皮における新規血管新生関連因子である

  山本和幸, 樋田泰浩, 加賀基知三, 平野聡, 樋田京子  日本外科学会雑誌  113-  (臨増2)  308  -308  2012/03  [Not refereed][Not invited]
  
• 腫瘍血管内皮におけるLysyl oxidaseの機能解析

  大澤崇宏, 樋田京子, 大賀則孝, 秋山廣輔, 樋田泰浩, 丸山覚, 安部崇重, 佐澤陽, 篠原信雄, 進藤正信, 野々村克也  日本泌尿器科学会雑誌  103-  (2)  356  -356  2012/03  [Not refereed][Not invited]
  
• 腫瘍微小環境におけるVEGFシグナルを介したMDR1の発現亢進による血管内皮細胞の薬剤抵抗性獲得について

  秋山廣輔, 大賀則孝, 樋田泰浩, 川本泰輔, 定本圭弘, 石川修平, 間石奈湖, 秋野文臣, 近藤美弥子, 松田彩, 井上農夫男, 進藤正信, 樋田京子  日本病理学会会誌  101-  (1)  287  -287  2012/03  [Not refereed][Not invited]
  
• Characteristics of tumor endothelial cells

  Kyoko Hida  Seikagaku  84-  (1)  43  -46  2012  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞におけるALDHの発現解析とALDH高活性血管内皮細胞の特性解析

  大村瞳, 大村瞳, 大賀則孝, 樋田泰浩, 秋山廣輔, 間石奈湖, 近藤美弥子, 大澤崇宏, 山本和幸, 川本泰輔, 飯田順一郎, 進藤正信, 樋田京子  日本がん転移学会学術集会・総会プログラム抄録集  21st-  91  2012  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞のがん転移への関わり

  間石奈湖, 大場雄介, 山本和幸, 大賀則孝, 浜田淳一, 秋山廣輔, 川本泰輔, 大澤崇宏, 近藤美弥子, 大村瞳, 樋田泰浩, 樋田京子  日本がん転移学会学術集会・総会プログラム抄録集  21st-  104  2012  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞の異常性

  HIDA Kyoko  生化学  84-  (1)  43  -46  2012  [Not refereed][Not invited]
  
• がん微小環境と腫瘍血管内皮細胞

  HIDA Kyoko  “Drug Delivery System”、 特集｢基礎から拓くDDS創薬フロンティア｣日本DDS学会  27-  (1)  34  -39  2012  [Refereed][Not invited]
   

  Tumor blood vessels play an important role in tumor progression and metastasis. We have shown that tumor endothelial cells (TEC) upregulate specific gene expressions. These molecules could be a novel target for for ideal antiangiogenic therapy.
• 腫瘍血管内皮細胞の異常と癌の悪性化との関連

  HIDA Kyoko  日本口腔腫瘍学会誌  24-  (3)  88  -94  2012  [Refereed][Not invited]
   

  Tumor blood vessels play an important role in tumor progression and metastasis. Thus, targeting the tumor blood vessels is an important strategy in cancer therapy. Tumor blood vessels generally arise from pre-existing vessels and have been thought to be genetically normal. However, they have been shown to differ from their normal counterparts e.g., with regard to morphological changes. We isolated tumor endothelial cells (TECs) from mouse tumor xenografts and showed that they were abnormal. TECs up-regulate many genes, proliferate more rapidly, and migrate more than normal endothelial cells (NECs). Furthermore, the TECs in our study were cytogenetically abnormal. TECs were drug resistant to paclitaxel with upregulation of multidrug resistance 1 (MDR1) mRNA, compared to NECs. Tumor-conditioned medium treatment caused drug resistance in NEC. We concluded that TECs can acquire cytogenetic abnormalities while in the tumor microenvironment.
  In order to develop ideal antiangiogenic therapies, it is important to understand the crosstalk between blood vessels and the tumor microenvironment.
  This review considers the current studies on TEC abnormalities, and discusses the possible mechanism by which the tumor microenvironment produces abnormal TECs.
• 低酸素環境が血管内皮細胞に及ぼす影響(Hypoxia is a key factor to acquisition of tumor endothelial cells abnormalities)

  近藤 美弥子, 大賀 則孝, 秋山 廣輔, 間石 奈湖, 川本 泰輔, 大澤 崇宏, 山本 和幸, 大村 瞳, 樋田 泰浩, 進藤 正信, 樋田 京子  日本癌学会総会記事  70回-  124  -124  2011/09  [Refereed][Not invited]
  
• 腫瘍血管内皮細胞とがん転移との相互作用解析(Analysis of interaction between tumor endothelial cells and tumor metastasis)

  間石 奈湖, 大賀 則孝, 樋田 泰浩, 大場 雄介, 浜田 淳一, 秋山 廣輔, 山本 和幸, 大澤 崇宏, 近藤 美弥子, 川本 泰輔, 進藤 正信, 井上 農夫男, 樋田 京子  日本癌学会総会記事  70回-  430  -430  2011/09  [Refereed][Not invited]
  
• 転移能の異なる腫瘍から分離された腫瘍血管内皮細胞の比較解析(Comparative characterization of tumor endothelial cells isolated from highly and low metastatic tumors)

  大賀 則孝, 石川 修平, 樋田 泰浩, 秋山 広輔, 間石 奈湖, 近藤 美弥子, 川本 泰輔, 進藤 正信, 樋田 京子  日本癌学会総会記事  70回-  430  -430  2011/09  [Refereed][Not invited]
  
• 腫瘍血管内皮におけるLysyl oxidaseの機能解析(The role of Lysyl oxidase on abnormal phenotypes of tumor endothelial cells)

  大澤 崇宏, 大賀 則孝, 北山 和子, 樋田 泰浩, 川本 泰輔, 近藤 美弥子, 間石 奈湖, 山本 和幸, 秋山 廣輔, 篠原 信雄, 野々村 克也, 進藤 正信, 樋田 京子  日本癌学会総会記事  70回-  430  -430  2011/09  [Refereed][Not invited]
  
• 血管の多様性 組織発生から疾患におけるダイナミクス 腫瘍血管内皮の多様性と薬剤抵抗性(Cellular and functional diversity of the vasculature: health and disease Heterogeneity of Tumor Endothelium and Drug Resistance)

  樋田 京子, 大賀 則孝, 秋山 廣輔, 樋田 泰浩, 進藤 正信  日本細胞生物学会大会講演要旨集  63回-  94  -94  2011/05  [Refereed][Not invited]
  
• Analysis of interaction between tumor endothelial cells and tumor cells

  Nako Maishi, Noritaka Ohga, Yasuhiro Hida, Yusuke Ohba, Taisuke Kawamoto, Kosuke Akiyama, Kazuko Kitayama, Miyako Kondoh, Takahiro Osawa, Kazuyuki Yamamoto, Nobuo Inoue, Masanobu Shindoh, Kyoko Hida  CANCER RESEARCH  71-  2011/04  [Not refereed][Not invited]
  
• 転移能の異なる腫瘍由来の血管内皮細胞の特性解析

  大賀則孝, 石川修平, 樋田泰浩, 秋山広輔, 間石奈湖, 川本泰輔, 近藤美弥子, 進藤正信, 樋田京子  日本臨床口腔病理学会総会・学術大会プログラム・抄録集  22nd-  145  2011  [Not refereed][Not invited]
  
• 緑茶ポリフェノールは腫瘍血管内皮に直接的な抑制効果を有する

  大賀則孝, 村木力, 黒須拓郎, 樋田泰浩, 土屋邦彦, 戸塚靖則, 進藤正信, 樋田京子  日本口腔科学会雑誌  60-  (1)  27  -27  2011/01  [Not refereed][Not invited]
  
• がん微小環境内の血管内皮の異常性

  樋田京子, 大賀則孝, 樋田泰浩, 進藤正信  日本病理学会会誌  99-  (2)  17  -17  2010/09  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞と腫瘍細胞との相互作用解析(Analysis of interaction between tumor endothelial cells and tumor cells)

  間石 奈湖, 大賀 則孝, 秋山 廣輔, 北山 和子, 近藤 美弥子, 川本 泰輔, 大澤 崇宏, 山本 和幸, 樋田 泰浩, 大場 雄介, 進藤 正信, 樋田 京子  日本癌学会総会記事  69回-  103  -103  2010/08  [Refereed][Not invited]
  
• 癌細胞培養上清による正常血管内皮の薬剤抵抗性の獲得(Emdothelial cells acquire drug resistance by factors from tumor cells)

  秋山 廣輔, 大賀 則孝, 樋田 泰浩, 黒須 拓郎, 北山 和子, 石川 修平, 近藤 美弥子, 間石 奈湖, 川本 泰介, 大澤 崇宏, 山本 和幸, 進藤 正信, 樋田 京子  日本癌学会総会記事  69回-  103  -103  2010/08  [Refereed][Not invited]
  
• がん患者末梢循環血管内皮細胞における遺伝子発現解析(Gene expression analysis of circulating endothelial cells in cancer patients)

  近藤 美弥子, 大賀 則孝, 黒須 拓郎, 北山 和子, 秋山 廣輔, 間石 奈湖, 川本 泰輔, 大澤 崇宏, 山本 和幸, 樋田 泰浩, 進藤 正信, 樋田 京子  日本癌学会総会記事  69回-  130  -131  2010/08  [Refereed][Not invited]
  
• 転移能の異なる腫瘍から分離された腫瘍血管内皮細胞の特性解析(Comparative characterization of tumor endothelial cells isolated from highly and low metastatic tumors)

  大賀 則孝, 石川 修平, 樋田 泰浩, 秋山 広輔, 北山 和子, 近藤 美弥子, 間石 奈湖, 川本 泰輔, 大澤 崇宏, 山本 和幸, 進藤 正信, 樋田 京子  日本癌学会総会記事  69回-  270  -270  2010/08  [Refereed][Not invited]
  
• 癌が分泌する小胞は内皮細胞の遺伝子発現を変化させる(Tumor-derived microvesicles cause gene changes in endothelial cells)

  川本 泰輔, 大賀 則孝, 北山 和子, 秋山 廣輔, 近藤 美弥子, 間石 奈湖, 大澤 崇宏, 山本 和幸, 樋田 泰浩, 進藤 正信, 樋田 京子  日本癌学会総会記事  69回-  497  -498  2010/08  [Refereed][Not invited]
  
• Adrenomedullin antagonist suppresses tumor formation in renal cell carcinoma through inhibitory effects on tumor endothelial cells and endothelial progenitor mobilization.

  Tsuchiya Kunihiko, Hida Kyoko, Hida Yasuhiro, Muraki Chikara, Ohga Noritaka, Akino Tomoshige, Kondo Takeshi, Miseki Tetsuya, Nakagawa Koji, Shindoh Masanobu, Harabayashi Toru, Shinohara Nobuo, Nonomura Katsuya, Kobayashi Masanobu  International journal of oncology  36-  (6)  1379  -1386  2010/06  

  Adrenomedullin (AM) is a multifunctional 52-amino acid peptide. AM has several effects and acts as a growth factor in several types of cancer cells. Our previous study revealed that an AM antagonist (AMA) suppressed the growth of pancreatic tumors in mice, although its mechanism was not elucidated. In this study, we constructed an AMA expression vector and used it to treat renal cell carcinoma (RCC) in mice. This AMA expression vector significantly reduced tumor growth in mice. In addition, microvessel density was decreased in AMA-treated tumors. To analyze the effect of AMA on tumor angiogenesis in this model, tumor endothelial cells (TECs) were isolated from RCC xenografts. TEC proliferation was stimulated by AM and it was inhibited by AMA significantly. AM induced migration of TECs and it was also blocked by AMA. However, normal ECs (NECs) were not affected by either AM or AMA. These results demonstrate that AMA has inhibitory effects on TECs specifically, not on NEC, thereby inhibiting tumor angiogenesis. Furthermore, we showed that vascular endothelial growth factor-induced mobilization of endothelial progenitor cell (EPC) into circulation was inhibited by AMA. These results suggest that AMA can be considered a good anti-angiogenic reagent that selectively targets TECs and EPC in renal cancer.
• PP-551 転移能の異なる腫瘍由来の血管内皮細胞の特性解析(発表・討論,一般演題ポスター,第98回日本泌尿器科学総会)

  石川 修平, 大賀 則孝, 秋野 文臣, 樋田 泰浩, 黒須 拓郎, 秋山 廣輔, 近藤 美弥子, 間石 奈湖, 篠原 信雄, 野々村 克也, 進藤 正信, 樋田 京子  日本泌尿器科學會雜誌  101-  (2)  506  -506  2010/02/20  [Not refereed][Not invited]
  
• 転移能の異なる腫瘍から分離された腫瘍血管内皮細胞の特性解析(Comparative characterization of tumor endothelial cells isolated from high and low metastatic tumors)

  石川 修平, 大賀 則孝, 樋田 泰浩, 秋野 文臣, 黒須 拓郎, 秋山 廣輔, 近藤 美弥子, 間石 奈湖, 野々村 克也, 進藤 正信, 樋田 京子  日本癌学会総会記事  68回-  212  -212  2009/08  [Refereed][Not invited]
  
• 緑茶ポリフェノールEGCGは腫瘍血管内皮に対する抑制効果を有する(Inhibitory Effects of Epigallocatechin-3 Gallate, a Polyphenol in Green Tea, on Tumor-Associated Endothelial Cells)

  大賀 則孝, 樋田 泰浩, 土屋 邦彦, 村木 力, 秋山 広輔, 近藤 美弥子, 間石 奈湖, 黒須 拓郎, 大廣 洋一, 戸塚 靖則, 進藤 正信, 樋田 京子  日本癌学会総会記事  68回-  219  -219  2009/08  [Refereed][Not invited]
  
• 癌細胞培養上清による正常血管内皮の異常性獲得(Endothelial cells acquire abnormalities by factors from tumor cells)

  秋山 廣輔, 大賀 則孝, 樋田 泰浩, 秋野 文臣, 黒須 拓郎, 石川 修平, 近藤 美弥子, 間石 奈湖, 井上 農夫男, 進藤 正信, 樋田 京子  日本癌学会総会記事  68回-  269  -269  2009/08  [Refereed][Not invited]
  
• 緑茶カテキンEGCGは腫瘍血管内皮細胞とならびに血管内皮前駆細胞に直接的な抑制効果を有する

  大賀則孝, 樋田京子, 樋田泰浩, 村木力, 土屋邦彦, 大廣洋一, 戸塚靖則, 進藤正信  日本病理学会会誌  98-  (1)  370  -370  2009/03  [Not refereed][Not invited]
  
• APP-003-AM 腎がん血管内皮細胞の染色体異常(総会賞応募ポスター,第97回日本泌尿器科学会総会)

  秋野 文臣, 樋田 京子, 樋田 泰浩, 土屋 邦彦, 大賀 則孝, 松田 光平, 黒須 拓郎, 石川 修平, 秋山 広輔, 佐澤 陽, 篠原 信雄, 進藤 正信, 野々村 克也  日本泌尿器科學會雜誌  100-  (2)  126  -126  2009/02/20  [Not refereed][Not invited]
  
• 癌と血管新生 5.癌組織中の血管内皮細胞の異常性―遺伝子発現から染色体異常まで

  樋田京子, 秋野文臣, 樋田泰浩  実験医学  27-  (2)  277-283,148(4)  2009/02/01  [Not refereed][Not invited]
  
• 緑茶ポリフェノールEGCGは腫瘍血管内皮に対する抑制効果を有する

  大賀則孝, 樋田泰浩, 土屋邦彦, 土屋邦彦, 村木力, 秋山広輔, 近藤美弥子, 間石奈湖, 黒須拓郎, 戸塚靖則, 進藤正信, 樋田京子  日本血管生物医学会  17th-  74  2009  [Not refereed][Not invited]
  
• がん組織中の血管内皮細胞の異常性－遺伝子発現から染色体異常まで

  HIDA Kyoko  実験医学増刊号「癌の微小環境と適応応答の最前線」  27-  (2)  149  -155  2009  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞の特性と治療抵抗性

  HIDA Kyoko  血管医学 特集「がんと血管－イメージングから治療まで－」  10-  (4)  23  -28  2009  [Not refereed][Not invited]
  
• 血管新生up‐to‐date 腫瘍血管内皮細胞の特性

  樋田京子, 樋田泰浩  細胞  40-  (12)  512  -515  2008/11  [Not refereed][Not invited]
   

  "腫瘍血管内皮は正常細胞である"という概念のもとに、正常血管内皮細胞を用いた研究の知見から腫瘍血管新生阻害剤が開発されてきた。しかし近年、腫瘍血管内皮細胞と正常内皮細胞の違いが知られるようになり、腫瘍血管内皮細胞の生態を明らかにすることが腫瘍血管により特異的に作用する血管新生阻害剤の開発につながるものと考えられる。われわれはこれまで腫瘍血管内皮細胞の分離培養を行い、特異遺伝子の発現や、抗癌剤への抵抗性を見いだしてきた。さらに腫瘍血管内皮細胞には染色体異常が認められた。腫瘍血管内皮細胞を用いた研究は腫瘍血管新生のメカニズムの解明に寄与するものと期待される。本章では腫瘍血管内皮細胞の異常性について概説する。(著者抄録)
• 腫瘍血管内皮細胞ならびに血管内皮前駆細胞は正常血管内皮細胞に比べ、緑茶ポリフェノールに感受性を有する(Tumor endothelial cells and EPC are more sensitive to EGCG compared to normal endothelial cells)

  大賀 則孝, 樋田 京子, 村木 力, 樋田 泰浩, 土屋 邦彦, 大廣 洋一, 戸塚 靖則, 進藤 正信  日本癌学会総会記事  67回-  365  -365  2008/09  [Refereed][Not invited]
  
• がん微小環境における腫瘍血管内皮細胞の異常性

  樋田京子, 樋田泰浩, 秋野文臣, 大賀則孝, 松田光平, 黒須拓郎, 土屋邦彦, 村木力, 進藤正信  補体シンポジウム講演集  45th-  178  2008/07  [Not refereed][Not invited]
  
• Neuropilins as Common Targeting Molecules on Tumor and Endothelial Cells to Inhibit Metastasis

  HIDA YASUHIRO, HIDA KYOKO, BIELENBERG DIANE, KLAGSBRUN MICHAEL, KONDO SATOSHI  Biotherapy (Tokyo)  22-  (2)  80  -86  2008/03/30  [Not refereed][Not invited]
   

  クラス3セマフォリン(SEMA3)は、神経ガイダンスと血管新生のシグナルを伝えるニューロピリン(NRP)のリガンドである。SEMA3とNRPは、当初神経細胞の軸索伸長の制御因子として見いだされたが、血管系や腫瘍生物学にも重要な役割を担うことが明らかになってきた。SEMA3は神経細胞を反発させて軸索成長円錐を退縮させるだけではなく、血管内皮細胞や腫瘍細胞に対しても同様の効果を示す。動物実験の結果、NRPは転移や血管新生の治療標的の候補であることが示された。また、異種移植動物実験では抗NRP-1抗体は抗VEGF抗体の抗腫瘍効果を増強した。NRP-2に結合するSEMA3Fは、腫瘍細胞の遊走と血管新生を阻害する。本稿では、腫瘍血管新生と転移におけるNRPの役割を細胞とマウスの実験結果から概説する。(著者抄録)
• Understanding tumor endothelial cell abnormalities to develop ideal anti-angiogenic therapies

  HIDA Kyoko, HIDA Yasuhiro, SHINDOH Masanobu  Cancer Sci  99-  (3)  459  -466  2008/03/10
• 口腔扁平上皮がんにおけるPim-1の発現は細胞運動能を亢進し転移活性に関与する

  田中 宗一, 北村 哲也, 樋田 京子, 東野 史裕, 戸塚 靖則, 進藤 正信  日本病理学会会誌  97-  (1)  221  -221  2008/03
• 腫瘍血管内皮細胞における特異マーカーの解析

  黒須拓郎, 樋田京子, 樋田泰浩, 藤江学, 小野寺雄一郎, 土屋邦彦, 村木力, 大賀則孝, 松田光平, 秋野文臣, 戸塚靖則, 進藤正信  日本病理学会会誌  97-  (1)  239  -239  2008/03  [Not refereed][Not invited]
  
• Targeting Endothelial and Tumor Cells with Natural VEGF Antagonist SEMA3F

  HIDA YASUHIRO, HIDA KYOKO, BIELENBERG DIANE, KLAGSBRUN MICHAEL, KONDO SATOSHI  癌の臨床  54-  (3)  157  -162  2008/03  [Not refereed][Not invited]
  
• 腎癌血管内皮細胞のaneuploidy

  秋野文臣, 樋田京子, 土屋邦彦, 樋田泰浩, 村木力, 大賀則孝, 松田光平, 黒須拓郎, KLAGSBRUN Michael, 原林透, 篠原信雄, 野々村克也, 進藤正信  日本病理学会会誌  97-  (1)  224  -224  2008/03  [Not refereed][Not invited]
  
• 腎がん血管内皮細胞のaneuploidy

  秋野文臣, 樋田京子, 土屋邦彦, 樋田泰浩, KLAGSBRUN Michael, 村木力, 大賀則孝, 松田光平, 原林透, 篠原信雄, 進藤正信, 野々村克也  日本泌尿器科学会雑誌  99-  (2)  251  -251  2008/02/20  [Not refereed][Not invited]
  
• 緑茶ポリフェノールによる腫瘍血管内皮細胞と血管内皮前駆細胞への抑制効果

  大賀則孝, 樋田京子, 樋田泰浩, 村木力, 土屋邦彦, 松田光平, 大廣洋一, 戸塚靖則, 進藤正信  生化学  1P-1394  2008  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞の特性

  HIDA Kyoko  細胞  40-  (12)  16  -19  2008  [Not refereed][Not invited]
  
• 腫瘍血管内皮を標的としたCyclooxigenase-2の血管新生阻害作用(Anti-Angiogenic Effects of Cyclooxygenase-2 inhibitor on Tumor-associated Endothelial Cells)

  村木 力, 樋田 京子, 樋田 泰浩, 土屋 邦彦, 戸塚 靖則, Klagsbrun Michael, 進藤 正信  日本癌学会総会記事  66回-  147  -147  2007/08  [Refereed][Not invited]
  
• 緑茶ポリフェノールによる腫瘍血管内皮と血管内皮前駆細胞への抑制効果(A green tea polyphenol, EGCG has inhibitory effects on tumor endothelial cells and endothelial progenitor cells)

  大賀 則孝, 樋田 京子, 村木 力, 樋田 泰浩, 土屋 邦彦, 戸塚 靖則, 進藤 正信  日本癌学会総会記事  66回-  523  -523  2007/08  [Refereed][Not invited]
  
• Natural VEGF antagonist,SEMA3Fによる腫瘍細胞と血管内皮細胞を標的とした転移抑制

  樋田泰浩, 樋田京子, BIELENBERG Diane, KLAGSBRUN Michael, 近藤哲  日本外科学会雑誌  108-  (臨増2)  76  -76  2007/03  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞の分離ならびに特異性の解析

  松田光平, 樋田京子, 土屋邦彦, 村木力, 樋田泰浩, 大賀則孝, 秋野文臣, 戸塚靖則, 進藤正信  日本病理学会会誌  96-  (1)  185  2007/02/05  [Not refereed][Not invited]
  
• Ets‐1がん遺伝子による口腔がん発生・悪性化機構―GnT‐Vの関与―

  喜屋武麻記子, 喜屋武麻記子, 小野寺雄一郎, 飯塚正, 樋田京子, 東野史裕, 北川善政, 進藤正信  日本口腔科学会雑誌  56-  (1)  118  -119  2007/01/10  [Not refereed][Not invited]
  
• がん微小環境における腫瘍血管内皮細胞の異常性

  樋田京子, 樋田泰浩, 進藤正信  生化学  4W12-4  2007  [Not refereed][Not invited]
  
• リンパ管発生/新生と腫瘍血管新生 5.腫瘍血管内皮細胞の特異性―理想的な血管新生阻害剤の開発をめざして

  樋田京子, 樋田泰浩  実験医学  24-  (18)  2852  -2857  2006/11/10  [Not refereed][Not invited]
  
• Cyclooxygenase-2 inhibitorは腫瘍血管新生を抑制する

  村木 力, 樋田 京子, 樋田 泰浩, 土屋 邦彦, 戸塚 靖則, Michael Klagsbrun, 進藤 正信  日本癌学会総会記事  65回-  312  -312  2006/09  [Refereed][Not invited]
  
• 緑茶ポリフェノールによる腫瘍血管内皮への抑制効果

  樋田京子, 村木力, 樋田泰浩, 土屋邦彦, 松田光平, KLAGSBRUN Michael, 進藤正信  日本癌学会学術総会記事  65th-  33  2006/08/28  [Not refereed][Not invited]
  
• 腫よう血管内皮の特異的性質 染色体遺伝子学的異常について

  樋田京子, 樋田泰浩, 土屋邦彦, 村木力, 小林正伸, KLAGSBRUN Michael, 進藤正信  日本病理学会会誌  95-  (1)  241  2006/03/18  [Not refereed][Not invited]
  
• 腫よう血管内皮におけるCyclooxygenase‐2 inhibitorの作用効果

  村木力, 樋田京子, 樋田泰浩, 土屋邦彦, 戸塚靖則, KLAGSBRUN Michael, 進藤正信  日本病理学会会誌  95-  (1)  241  2006/03/18  [Not refereed][Not invited]
  
• Tumor-associated Endothelial Cells as Targets of Anti-angiogenic Therapy

  Klagsbrun Michael  Journal of Japan Surgical Society  107-  (2)  2006/03/05  [Not refereed][Not invited]
  
• 腫瘍血管内皮細胞の特異性－理想的な血管新生阻害剤の開発を目指して

  HIDA Kyoko  総説 実験医学増刊号「急速進展する血管研究」  24-  (18)  152  -157  2006  [Not refereed][Not invited]
  
• 腫よう関連血管内皮細胞は細胞発生学的に異常を伴っている

  樋田京子, 樋田泰浩, 進藤正信, KLAGSBRUN Michael  日本癌学会学術総会記事  64th-  43  2005/08/15  [Not refereed][Not invited]
  
• セマフォリン3Fは悪性黒色腫の転移,血管新生,および血管内皮前駆細胞の動員を抑制する

  樋田泰浩, 樋田京子, 清水昭夫, BIELENBERG Diane, MOSES Marsha, KLAGSBRUN Michael  日本癌学会学術総会記事  64th-  44  2005/08/15  [Not refereed][Not invited]
  
• HPCフィルムの口腔内被覆材としての応用

  笠師 久美子, 原田 幸子, 和田 育男, 小林 一三, 船岡 孝誠, 樋田 京子, 戸塚 靖則  歯科薬物療法  18-  (3)  110  -115  1999  [Refereed][Not invited]
  
• MRI evaluation of the effect of stabilization splints in patients with TMJ closed lock

  YURA Shinya, TOTSUKA Yasunori, KUDO Yoshifumi, HORIMUKAI Hiromasa, SUZUKI Toyonori, YOSHIKAWA Tetsuya, YASUDA Mutsumi, HIDA Kyoko, KADOWAKI Shigeru, KOBAYASHI Takashi, YAMAGUCHI Taihiko, INOUE Nobuo  Journal of Oral Surgery Society of Japan  44-  (10)  787  -793  1998  [Refereed][Not invited]
   

  Fourteen patients (17 joints) with TMJ closed lock treated by stabilization splints, were evaluated with regard to condyle movement and changes in the disc and bone by means of MR imaging before and after treatment.
  In all 13 joints that responded to treatment, condyle mobility increased after treatment. Disc displacement and deformity progressed in 3 joints. Disc reduction was seen in 2 joints, and although 11 joints were still locked, disc mobility improved in association with structural changes. In 4 joints with no response to treatment, these changes were not seen. Pseudodisk formation and osseous changes were not observed in any patient.
  These results suggest that the use of stabilization splints in patients with closed lock may promote disc reduction and increase condylar and disc mobility even when the disc is still locked.
• Expression of E1AF, an ets-family transcription factor, is correlated with the invasive phenotype of oral squamous cell carcinoma

  K Hida, M Shindoh, K Yoshida, A Kudoh, K Furaoka, T Kohgo, K Fujinaga, Y Totsuka  ORAL ONCOLOGY  33-  (6)  426  -430  1997/11  [Refereed][Not invited]
   

  E1AF is a newly identified ets-oncogene family transcription factor. Previous reports have noted that E1AF can upregulate promoter activities of several matrix metalloproteinase (MMP) genes and showed that invasive potentials of oral squamous cell carcinoma-derived cell lines are correlated with expression of E1AF and MMPs. The invasive phenotype is restrained by transfection with an antisense E1AF expression vector. Thus, E1AF is thought to be highly correlated with malignant potentials of cancer cells. However, little is known about E1AF expression and cancer cell malignancies in in vivo tumours. In the present study, 27 oral squamous cell carcinoma (SCC) specimens were examined using RT-PCR, Southern blot hybridisation and in situ hybridisation (ISH) and compared to the clinicopathological parameters. Among the 27 patients, E1AF was detected in 15 cases. E1AF mRNA was detected in 13 of 17 invasive SCCs, whereas the majority of SCCs not expressing E1AF showed an expansive growth pattern. Increased prevalence of E1AF-positive oral SCC was observed in cases with nodal metastasis. These results indicate that E1AF may be involved in cancer cell malignancies through its ability to promote invasive potential. (C) 1997 Elsevier Science Ltd. All rights reserved.
• Application of pumping manipulation technique to patients with persistent TMJ closed lock : Correlation between arthrographic findings and therapeutic results

  HIDA Kyoko, YURA Shinya, ANAZAWA Toshiyuki, SATOH Jun, SATOH Chiharu, KOBAYASHI Takashi, INOUE Nobuo, TOTSUKA Yasunori  TMJ : journal of Japanese Society for Temporomandibular Joint : 日本顎関節学会雑誌  9-  (1)  130  -138  1997/06/20
• 慢性クローズド,ロック症例に対する顎関節パンピング,マニピュレーション法の応用ー造影所見と治療効果との比較

  由良晋也, 戸塚靖則, 工藤善史, 堀向弘眞, 鈴木豊典, 由川哲也, 安田 睦, 樋田京子, 門脇 繁, 小林 隆, 山口泰彦, 井上農夫男  日顎誌  (9)  129  -137  1997  [Refereed][Not invited]
  
• Clinical history and morphological changes of temporomandibular joint structures in patients with anterior disk displacement without reduction

  KADOWAKI Shigeru, NAKAJIMA Yoritoshi, DOIGAMI Teruo, WATANABE Masaki, KOBAYASHI Takashi, YURA Shinya, HIDA Kyoko, UEDA Michihiro, MURAKAMI Yuji, INOUE Nobuo, TOTSUKA Yasunori  TMJ : journal of Japanese Society for Temporomandibular Joint : 日本顎関節学会雑誌  8-  (3)  495  -506  1996/12/20
• パラフィン標本からのPCR (PCR法最前線--基礎技術から応用まで) -- (PCRの反応組成と条件)

  進藤 正信, 樋田 京子, 船岡 孝誠  蛋白質核酸酵素  41-  (5)  463  -466  1996/04
• 非復位性の関節円板前方転位症例における臨床経過と顎関節構成体の形態変化

  門脇 繁, 中島頼俊, 土井上輝夫, 渡辺政明, 小林 隆, 由良晋也, 樋田京子, 上田倫弘, 村上有二, 井上農夫男, 戸塚靖則  日顎誌  (8)  21  -32  1996  [Refereed][Not invited]
  
• Therapeutic result of lavage manipuration technique.

  YURA Shinya, TOTSUKA Yasunori, HIDA Kyoko, KADOWAKI Shigeru, KOBAYASHI Takashi, TSUYAMA Masashi, YAMAGUCHI Taihiko, INOUE Nobuo, HARADA Hiroyuki, OHRUI Shin, KURODA Mitsugu  Journal of Oral Surgery Society of Japan  40-  (4)  519  -524  1994  [Refereed][Not invited]
   

  To evaluate the efficacy of lavage manipulation technique, 15 patients with persistent closed lock of the TMJ were treated by this method. The patients consisted of 1 male and 14 females. Age of the patients ranged from 18 to 66 years, with an average age of 33 years.
  This technique was followed by pumping of the upper compartment to distend the joint space after local anesthesia with sedation in the outpatient clinic. Two 18-gauge needles were inserted into the superior compartment to enable a free flow of the solution through the joint space. Physiological saline solution was injected via one of the needles under a sufficient pressure to irrigate 200 to 500 ml of the solution a during 20-minute period. During a 3-month follow-up of the method, interincisal mouth opening of the patients improved from a range of 7 to 25 mm, with an average opening of 14.9 mm.
  The results indicate that lavage manipulation technique is an effective method for treating patients with closed lock which can be performed under local anesthesia with sedation in the outpatient clinic.
• Comparative study of tomographic arthrographic findings and arthroscopic findings.

  YURA Shinya, TOTSUKA Yasunori, HIDA Kyoko, SATOH Jun, TSUYAMA Masashi, KADOWAKI Shigeru, YAMAGUCHI Taihiko, NAKAMURA Takeyuki, INOUE Nobuo, FUKUDA Hiroshi, MIURA Hisanori, NAKAMURA Hiroyuki  Journal of Oral Surgery Society of Japan  39-  (10)  1049  -1056  1994  [Refereed][Not invited]
   

  Although radiographic examination is generally used to diagnose lesions in the temporomandibular joint (TMJ), its reliability is still controversial. In this study, radio tomographic and single-contrast arthro tomographic findings were compared with arthroscopic findings in 34 joints of 28 patients with TMJ closed lock. The patients included 6 males and 22 females. Their ages ranged from 15 to 69 years old, with an average of 33 years.
  With regard to disk displacement, arthrographic findings agreed with arthroscopic findings in 33 out of 34 joints (97%). Regarding disk movement, both findings coincided in 29 out of 30 joints (97%). On the other hand, concidence of arthrographic and arthroscopic findings in disk deformation, anterior joint compartment adhesion and disk or retrodiscal tissue perforation were 53%, 66% and 74%, respectively. Moreover, tomographic and arthrographic examinations did not show abnormality on the surface of the mandibular fossa or articular eminence.
  These results suggest that tomography and single-contrast arthrography do not necessarily provide high reliability in the assessment of intracapsular lesions of the TMJ.

Awards & Honors

• 2024/05 歯科基礎医学会 第24回歯科基礎医学会ライオン学術賞
   がん関連血栓症における腫瘍血管内皮細胞の役割の解明
• 2023/10 国立大学法人北海道大学 令和5年度国立大学法人北海道大学桂田芳枝賞
   

  受賞者: 樋田京子
• 2018/05 資生堂 第11回資生堂女性研究者サイエンスグラント
   高転移性腫瘍細胞外小胞由来miRNAは腫瘍血管内皮における 薬剤耐性を誘導する 

  受賞者: 樋田京子
• 2017/03 The Society of Japanese Women Scientists The Society of Japanese Women Scientists Encouragement Prize
   新しいがん治療法の開発を目指したがん微小環境における血管の異常性解明 

  受賞者: HIDA Kyoko
• 2012/12 The 10th Korea-Japan Joint Symposium on Vascular Biology The 20th annual Meeting of the Japanese Vascular Biology and MedicineOrganization,
   Characterization of stem-like tumor endothelial cells 

  受賞者: Kyoko HIDA
• 2010 平成17年度 日本病理学会学術研究賞
  
• 2005 平成17年度 湯浅記念財団賞
  
• 2005 第50回日本口腔外科学会 メダルティス賞受賞
  

Research Grants & Projects

• A drug development study for radiation osteonecrosis of jaw with improving micro vascular-skeletal structure

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2023/06 -2025/03 

  Author : 飯村 忠浩, 丸岡 豊, 樋田 京子, 李 智媛
• Development of new diagnostic method of bladder cancer based on mRNA derived from urine exosome

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2022/04 -2025/03 

  Author : 大澤 崇宏, 伊藤 陽一, 安部 崇重, 樋田 京子, 篠原 信雄
• がんと感染症に共通した血栓形成機序の解明

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Research (Exploratory)

  Date (from‐to) : 2022/06 -2024/03 

  Author : 樋田 京子, 澤 洋文
• がんと腫瘍血管の相互作用による新たな転移促進機構の解明とその制御

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2021/04 -2024/03 

  Author : 菊地 奈湖, 樋田 京子, 樋田 泰浩, 篠原 信雄, 大廣 洋一, 松田 彩

   

  転移は予後を左右する重要な因子であり，がん治療において転移の予測やその制御は重要である．申請者らはこれまで腫瘍血管を裏打ちする腫瘍血管内皮細胞の異常性について検討し，腫瘍血管内皮細胞由来液性因子によりがんの転移が促進することを報告した．したがって，腫瘍血管内皮細胞は単なる血行性転移の経路の入り口となるだけではなく，積極的にがんの悪性化にも影響を与えうることが示唆された．がんの転移は，原発巣での浸潤ならびに血管内侵入，血管内の移動，転移先臓器への漂着，転移先臓器での増殖・転移巣形成という複雑なカスケードを経て成立する．異常性を示す腫瘍血管内皮細胞は，転移の初期段階だけではなく，さまざまな過程において関与する可能性があるのではないかと考えた．最近，がん組織内で腫瘍血管内皮細胞ががん細胞と細胞塊を形成することが示唆されている．実際，手術摘出がん組織標本において，血管腔内に血管内皮細胞で囲まれる細胞塊が観察された．そこで本研究では，血液中で腫瘍血管内皮細胞ががん細胞と細胞塊を形成し，転移促進などがんの悪性化に関与すると仮説を立て，腫瘍血管内皮による新たながん悪性化促進機構を明らかにすることを目的とした．手術摘出組織標本を用いて，血管腔内における細胞塊をH-E染色ならびに血管内皮マーカーCD31の組織免疫染色で可視化し，評価した．腫瘍組織の血管腔内に血管内皮細胞で構成される細胞塊が存在することが示唆された．がん患者血液中に細胞塊が存在するかどうか，ショ糖密度勾配遠心法で単核球分画を濃縮し，赤血球を除いて検討したところ，UEA-1レクチンで染色される血管内皮細胞を含む細胞塊が検出された．さらに，Poly-HEMA coatingした非接着カルチャープレート上でがん細胞と血管内皮細胞を共培養して細胞塊を形成後，そのphenotypeについて検討した．
• 腫瘍血管トランスポーターを標的とした薬剤耐性口腔がんの制御

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2021/04 -2024/03 

  Author : 梅山 悠伊, 樋田 京子, 菊地 奈湖, 松田 彩
• Development of new anti-cancer strategy via activation of immunity by targeting tumor endothelial factors

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2021/04 -2024/03 

  Author : 樋田 京子, 菊地 奈湖, 樋田 泰浩, 大廣 洋一, 松田 彩, 上田 倫弘
• 腫瘍特異的血管新生阻害剤とドラッグリポジショニングによる新規肺癌治療の開発

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2021/04 -2024/03 

  Author : 樋田 泰浩, 菊地 奈湖, 樋田 京子
• Development of oncolytic virus targeting tumor angiogenesis factor

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2020/04 -2023/03 

  Author : 松田 彩, 菊地 奈湖, 樋田 京子, 東野 史裕

   

  アデノウイルス（Ad）の初期遺伝子のタンパクはARE-mRNAを核外輸送・安定化する機能を有する。申請者らはこの機能がAdの複製に必須であることを見いだし、腫瘍溶解ウイルス（AdΔE4）を開発した。腫瘍血管は腫瘍組織へ栄養や酸素を供給し、がんの進展や転移に重要な役割を果たしていることが知られている。申請者らは腫瘍血管内皮のマーカーを同定し、腫瘍血管内皮が分泌するタンパクが腫瘍の進展や転移にどのように影響するかを解明してきた。腫瘍間質ではARE-mRNAが安定化されており、AdΔE4が増殖しやすい環境であることが予測される。そこで腫瘍血管内皮マーカーの存在下でわれわれの開発した腫瘍溶解ウイルスの効果が増強されるかを検討することとした。 これまでの研究で腫瘍血管が分泌するBiglycan (BGN)が腫瘍溶解ウイルスの効果を増強させることが示唆されている。 そこでBGNが担癌マウスモデルにおいても同様に腫瘍溶解ウイルスの効果を増強するか検討することとした。ヌードマウスの皮下にヒトがん細胞を移植し、形成した腫瘍に腫瘍溶解ウイルスを直接腫瘍内投与した。その結果、コントロールと比較し、腫瘍溶解ウイルスを投与した群では腫瘍のサイズが有意に小さく、腫瘍溶解効果が示された。マウスから腫瘍をサンプリングし、組織免疫染色により、癌細胞におけるアデノウイルスとHuRの発現を検討したところ、アデノウイルスが検出された細胞ではHuRが細胞質に発現している傾向がみられた。また腫瘍血管内皮細胞にBGNの発現がみられ、またHuRが腫瘍血管内皮細胞の細胞質に発現していることが示された。この結果より腫瘍溶解ウイルスは腫瘍細胞および腫瘍血管内皮細胞に効果があることが示唆された。
• 血管の制御による重症化COVID-19治療薬の開発

  橋渡し研究プログラム2022年度研究開発シーズA：

  Date (from‐to) : 2022/04 -2023/03
• Exploratory research of innovative pulp capping materials both with promoting healing of dental pulp and adhesive property to hard tissue

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2019/04 -2022/03 

  Author : sano hidehiko

   

  Based on the study by Toida et al, (Dent Mater J. ;41:126-133 2022), the combination of MTA and phosphorylated-pullulan resulted in good healing of exposed monkey pulp. Moreover, adhesive property of phosphorylated-pullulan made it possible to user friendly handlings and perfect dentin sealing. A report of pupal reactions after pulp capping treatments from Islam R et al, (Int Endod J. ;54:1902-1914 2021) stated interesting results. They used 4-META/MMA-TBB resin as a control material. The combination of MTA and phosphorylated-pullulan showed superior healing of pulp after application, compared to plain use of MTA. Additionally, plain use of phosphorylated-pullulan to the pulp did not show any adverse effect on the pulp healing. Preliminary study using colloidal platinum nanoparticles (CPN) to the exposed pulp demonstrated excellent pulp healing with less inflammatory reactions and prompt angiogenesis within the exposed pulp which might contribute to support good healing of the pulp.
• エナメル上皮腫の浸潤機構解明による新規治療法の開発

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  Date (from‐to) : 2019/04 -2022/03 

  Author : 北村 哲也, 菊地 奈湖, 大廣 洋一, 樋田 京子, 東野 史裕, 松田 彩

   

  本研究ではエナメル上皮腫の浸潤性増殖の解明を目的とし、主にbudding領域について詳細な検討を行ってきた。budding領域は、エナメル上皮腫の組織学的特徴の一つで、腫瘍から間質への出芽様構造のことであり、これが浸潤性増殖の原因と考えられている。これまで我々は、budding領域にHuRタンパク質と⊿Np63タンパク質が高発現していることを免疫組織学的に明らかにしてきた。そこで、HuRと⊿Np63との分子生物学的関連について着目した。HuRはmRNA結合タンパクであることから、HuRタンパクと⊿Np63 mRNAとの結合をRIP assay法によって検討した。細胞はエナメル上皮腫の培養細胞株であるAM-1細胞を用いた。その結果、HuRタンパクは⊿Np63 mRNAに結合することが明らかとなった。また、HuRタンパクとmRNAの結合を特異的に阻害するCMLD2を用いたところ、HuRタンパクと⊿Np63 mRNAとの結合は阻害された。 次に、HuRタンパクと⊿Np63 mRNAの結合の意義について検討した。AM-1細胞にCMLD-2を用いて、⊿Np63 mRNAの発現をreal time PCR法を用いて、⊿N63タンパク質の発現についてWestern blotting法を用いて検討した。⊿Np63 mRNAの半減期はCMLDを作用させると短くなり、⊿N63タンパク質は経時的に発現が減少した。このことから、HuRタンパクは⊿Np63 mRNAに結合して、mRNAを安定化させ、その結果タンパク質の発現を上昇させていることが明らかとなった。 これらの結果から、budding領域での生物学的機序が明らかとなった。
• Association between oral bacteria and oral cancer metastasis

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2019/04 -2022/03 

  Author : 大廣 洋一, 北村 哲也, 樋田 京子, 長谷部 晃

   

  細菌特異的な16s rDNAの発現をPCRで確認する際，バックグラウンドの低下と，精度を向上させるために下記を行った． 1) 腫瘍組織選択的なDNA 抽出条件の検討：手術標本の残余組織を用いて，①未染のパラフィン固定された組織，②ヘマトキシリン・エオジン（HE）染色された切片よりNucleoSpin DNA FFPE XSにてDNAを抽出した．HE染色により，腫瘍組織より選択的にDNAを回収できることを想定したが，HE染色によりDNA収量が30％程度以下になることを確認した．HEで染色では腫瘍部・非腫瘍部を判別するには簡便だが，DNAの抽出には不利であることがわかった． 2）ポジティブコントロールの調整：手術残余組織と細菌学教室より供与いただいたストレプトコッカス・ミュータンス（S. mutans）とを混合し，DNA抽出，PCRを行った．また，同様に大腸菌（E. Coli）を材料として上記手順を行ったところ組織とS. mutansの混合物ならびにE. Coliから回収したサンプルでは16sｒDNAの発現が確認された． 3）転移リンパ節標本からのDNA抽出条件の検討：リンパ節標本（15ｘ15mm大）ならびに原発腫瘍の深部（細菌に汚染されていない部分）のみからなる薄切（4.25 μm）切片10枚よりDNAを抽出し，さらにポジティブコントロールとして大腸菌（E.Coli）由来のDNAを用いて，PCRを行ったところ30サイクルではポジティブコントロールのみで発現したが，実験群では発現は確認できなかった．PCRの条件に問題はないが，標的臓器で確認するためには，キットの選定，プライマーの設定，サイクル数などの検討が必要と考えられた．
• COVID-19重症化における血管内皮細胞の感染病態解明と治療法の開発

  日本医療研究開発機構 (AMED)：新興・再興感染症に対する革新的医薬品等開発推進研究事業

  Date (from‐to) : 2021/04 -2022/03 

  Author : 樋田京子, 澤, 洋文, 樋田泰浩, 間石奈湖
• high-throughput screening for tumor endothelial cell specific inhibitors

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2018/04 -2021/03 

  Author : Hida Yasuhiro

   

  Tumor endothelial cells from human clinical tumors and normal endothelial cells from non-cancerous portions were isolated and cultured. FACS analysis and RT-PCR confirmed the establishment of highly pure primary cultured cells. The hTERT gene was transfected into these primary cultured cells, and more than 50 generations of passages were possible. The transgenic cells maintained a endothelial cell phenotype. The transgenic cells retained the phenotype of endothelial cells and the characteristic differences in gene expression observed in primary cultured cells were also retained, and it was confirmed that the cells met the conditions for use in cell-based high through-put assays. We are validating the inhibitors found by screening and verifying in the preclinical models.
• Elucidation of the mechanism for cancer metastasis induced by oral cancer exosome and development of novel therapy for metastatic cancer

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2018/04 -2021/03 

  Author : Hida Kyoko

   

  Cancer actively undergoes angiogenesis due to its progression and metastasis, and exosomes secreted by cancer cells are attracting attention because they change the traits of surrounding stromal cells. In this study, we will identify the exosome miRNA of cancer that causes phenotypic changes in the blood vessels of the primary lesion and distant organs and is involved in metastasis, clarify its molecular mechanism, and lead to the construction of a cancer metastasis inhibition strategy. In an in vivo tumor model, we showed that cancer miRNA X is transported by exosomes and taken up by tumor vascular endothelial cells. We found that miRNA X attenuates the barrier function of the vascular endothelium and promotes metastasis, and clarified a new metastasis mechanism.
• Basic research for elucidation of the mechanisms of tumor metastasis by tumor endothelial cells

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2018/04 -2021/03 

  Author : Kikuchi (Maishi) Nako

   

  We previously reported that endothelial cells of tumor blood vessels contribute to tumor metastasis. Since it is known that tumor cells aggregate in blood, which are involved in tumor malignancy, we investigated the possibility that endothelial cells also contribute to form tumor aggregation in this study. Tumor cells and endothelial cells formed cell aggregations when they were co-cultured in non-adherent condition. We found that the characteristics of endothelial cells were involved in tumor cell survival and proliferation in the cell aggregations. After forming cell aggregations, we isolated tumor cells and endothelial cells respectively by flow cytometory and analyzed gene expression patterns and activated pathways by IPA.
• 腫瘍血管・がん微小環境のネットワークの解明と新規血管新生阻害療法の開発

  日本医療研究開発機構 (AMED)：革新的がん医療実用化研究事業-がんネットワークの臨床的意義の理解に基づく医療シーズの開発研究

  Date (from‐to) : 2018/05 -2021/03 

  Author : 樋田京子,間石奈湖,樋田泰浩,西原広史,田中伸哉,篠原信雄
• 細胞社会をつなぐ血管内皮細胞のダイバーシティー獲得機構の解明

  日本学術振興会：科学研究費助成事業 新学術領域研究(研究領域提案型)

  Date (from‐to) : 2018/04 -2020/03 

  Author : 樋田 京子

   

  Biglycanノックアウトマウスを用いてin vivo 腫瘍モデルを用いてがん間質細胞と血管の相互的な関係におけるbiglycan の機能について解析した。腫瘍血管が発現するBiglycanが癌関連マクロファージや細胞傷害性リンパ球の動員の抑制に働いている知見を見出した。このことは腫瘍内の血管内皮によっても、こうした分子を発現しているものとそうでないものがあり、多様な性質をもち、さらには周囲の免疫環境にも異なる影響を及ぼしている可能性が示唆された。また，腫瘍血管内皮細胞のBiglycan により周囲間質の線維化が促進されることがわかり，免疫細胞の癌組織内の遊走を抑制する要因の一つになっていることが示唆された． また、治療経過による環境変化と上皮EMTと血管内皮細胞の相互作用についても検討を行った。 一次化学療法としてゲムシタビン，シスプラチンにより治療された尿路上皮がん患者の病理検体を用いた解析において，抗癌剤治療によるがん細胞のIL-8などのサイトカイン量の増加により、腫瘍血管内皮細胞においてNF-kBが活性化し，薬剤耐性関連分子ABCB1 の発現レベルが上昇することを見出した．2次治療として用いられるパクリタキセルに対する耐性の原因の一つとなることをin vitro, in vivo 両方において証明した． 治療経過による微小環境の変化によりがん細胞と血管の間の相互作用も変化しうることが示唆された。特に血管内皮細胞において治療中に発現変化する分子はその後の癌細胞の血行転移にも関連するため，治療標的として重要であることが示唆された．
• Development of the drugs targeting lung tumor endothelial cells and the companion diagnosis

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2012/04 -2016/03 

  Author : Hida Yasuhiro, KAGA KICHIZO, MATSUI YOSHIRO

   

  Tumor endothelial cells (TEC) from mouse xenografted tumor, especially that from highly metastatic tumor expresses high level of biglycan. Knock down of biglycan by siRNA suppressed tumor cell invasion thorough endothelial cell sheet. Co-implantation of low metastatic tumor cells with TEC developed lung metastases. This metastatic ability was abandoned by suppression of biglycan expression in TEC. Immunohistochemistry of human clinical tumors revealed specific expression of biglycan in TEC. ELISA of serum showed high biglycan level in cancer patients, especially those with metastasis.
• Development of novel therapy targeting cancer stem cells by ROS regulation

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2012/04 -2015/03 

  Author : TOTSUKA yasunori, OHGA Noritaka, TEI Kanchu, OHIRO Yoichi

   

  Reactive Oxygen Species (ROS) are known as the origin of oxidative stress. ROS are produced in response to hypoxia and nutrient deprivation. It is reported that tumor microenvironment is often exposed to hypoxic condition and nutrient deprivation because of the insufficient blood flow. ROS are accumulated in various cells in tumor microenvironment, and they promote cancer malignancy. In this study, we analyzed the effect on cancer stem cells and tumor blood endothelial cell that is reported to play a role as stem cell niche. ROS accumulation caused in alteration of gene expression profile both in cancer cells and tumor endothelial cells. One secreted protein from tumor endothelial cells was upregulated by ROS and attracted cancer cells. We analyzed the effect of ROS inhibition on cancer stem and tumor angiogenesis.
• Development of the treatment that targeted an arachidonate cascade of the oral cancer neighborhood environment

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2012/04 -2015/03 

  Author : KOBORI YOSHINORI, OGA Noritaka

   

  We found that COX-2/ prostaglandin E2 (PGE2) and prostacyclin (PGI2) was expressed highly in a tumor endothelial cells.In others to use expensive molecular target medicine for as a treatment strategy of the vascularization inhibition therapy, the COX-2/ prostaglandin E2 (PGE2) inhibitor could used cancer treatment.Targeting tumor angiogenesis is an established strategy for cancer therapy.
• Investigation of the mechanism of enhanced lung metastasis due to exosome from tumor endothelial cells

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2012/04 -2014/03 

  Author : HIDA Yasuhiro, KAGA Kichizou, MATSUI Yoshirou, KATOH Tatsuya, HIDA Kyoko

   

  We hypothesized that tumor-associated endothelial cells (TEC) facilitate tumor metastasis. We established TEC from xenograft tumor of the low metastatic malignant melanoma cell , A375 and from that of high metastatic counter part, A375SM("super metasitatic"). Comparison of those 2 TEC revealed specific up regulation of some microRNA in A375SM TEC. Transfection of the microRNA change the phenotype of normal endothelial cells from normal skin to that of A375SM TEC.
• 口腔環境が口腔がんの悪性化・上皮間葉移行を誘導する機構の解明

  日本学術振興会：科学研究費助成事業 挑戦的萌芽研究

  Date (from‐to) : 2012/04 -2014/03 

  Author : 進藤 正信, 北村 哲也, 樋田 京子, 東野 史裕

   

  口腔がんの腫瘍周囲環境が腫瘍の発生、増殖、浸潤、転移等の悪性化に関与する因子を検索することを目的に研究を行った。副甲状腺 関連タンパクPTHrPは乳がんや前立腺癌の骨転移において重要な役割を果たすことが知られている。口腔がん細胞でもPTHrPが高発現していることを我々は見いだした。さらに実際の口腔がん患者でPTHrP陽性率と腫瘍の転移・再発に有意な相関があることも明らかにし た。腫瘍間質に存在する線維芽細胞は腫瘍血管と同様に正常細胞とは異なった形質を発現していることが報告され、cancer-associate d fibroblast (CAF)と称されている。口腔領域に発生する粘表皮がんで検索したところ、PTHrPを高発現する粘表皮がんの中間細胞集 にCAFが多くみられ、さらに新生血管が豊富に存在し、PTHrP発現腫瘍細胞の悪性形質との関連が示唆された。線維芽細胞株MRC5をPTHr P高発現細胞株HSC2の上清を加えたMRC5では明らかな増殖活性の亢進がみられた。PTHrPsiRNA導入HSC2細胞ではPTHrPのmRNA発現が低下 していることが示され、PTHrPノックダウンHSC2細胞培養上清を加えたMRC5細胞は増殖活性の減弱がみられた。さらに通常培養ではαS MAの発現がほとんどみられないMRC5、DP31、DP36をPTHrP産生癌細胞上清で培養したところ、線維芽細胞株でαSMAの発現が亢進することがWestern blotであきらかになり、PTHrPによる周囲間質に存在する線維芽細胞のがん関連線維芽細胞(CAF)への誘導効果が示された。
• 血管を標的とする革新的医薬分子送達法の基盤技術の確立

  文部科学省：特別教育研究経費（研究推進）

  Date (from‐to) : 2009/04 -2014/03 

  Author : 樋田 京子
• 腫瘍血管と微小環境との相互作用の解明とその分子機構を標的とした治療法開発

  日本学術振興会：科学研究費助成事業 新学術領域研究(研究領域提案型)

  Date (from‐to) : 2011/04 -2013/03 

  Author : 樋田 京子

   

  １）がんの培養上清による血管内皮のmiRNAの発現変化 がんの培養上清による血管内皮の遺伝子発現変化については，それががん由来のmiRNAや遺伝子の取り込みによるものなのか，血管内皮側の転写亢進によるものかなどを検討する．高転移性の腫瘍の培養上清中のエクソソームには低転移性の腫瘍放出エクソソームと異なったmiRNAXが含まれていることがわかった．このmiRNAXがホストの血管内皮にエクソソームを介して取り込まれるのかどうかについてしらべた．このmiRNAが確かにエクソソームを介して血管内皮に取り込まれること，さらにその発現が血管内皮においてあがることが確認できた．現在このmiRNAの血管内皮における機能の解析を行っている．２) 異常性を獲得する分子機構を標的とした治療 これまでの研究で腫瘍由来のVEGFにより血管内皮の膜トランスポーターP-gpの発現が上がることがわかった．これを阻害するverapamilによりin vitro の系では血管内皮の薬剤感受性が回復することがわかり，この薬剤を併用した抗がん剤低用量使用による血管新生阻害療法をおこなっている．３) がんの進展における経時的な腫瘍血管内皮細胞の遺伝子発現解析 北海道大学病院において肺がん，大腸がん，胃がん，腎がん，前立腺がんや口腔がんと診断された症例において，ヒト腫瘍病理検体や末梢血を用いてTECマーカーの発現を解析し，がんの悪性度との関連や治療経過とがん組織像，血管の成熟化との関連などを解析し現在，臨床パラメータとの比較をおこなった．現在論文作成中である．
• Discovery of diagonositc and therapeutic biomarker by circulating tumor endothelial cells in metastatic renal cell carcinoma

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2011 -2013 

  Author : SHINOHARA Nobuo, HIDA Kyoko, HIDA Yasuhiro, NONOMURA Katsuya

   

  We identified several markers (PTGIR, Biglycan, LOX, CXCR7 etc. ) highly expressed in mouse tumor endothelial cell (TEC) by real-time PCR of circulating endothelial cell (CEC) in metastatic renal cell carcinoma(RCC). We isolated human TEC from human RCC tissues and human normal endothelial cell (NEC) from normal kidney tissues of six patients. PTGIR, Biglycan, LOX and CXCR7 expression levels were significantly higher in human TEC than in human NEC for all paired cases. These results suggested that CEC might be a useful biomarker of RCC.
• Development of novel therapies by antioxidants for reactive oxidative species of tumor microenvironment in oral carcinoma

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2011 -2013 

  Author : MATSUZAWA Yusuke, OGA Noritaka, HIDA Kyoko

   

  It is known that inflammatory cells pruduce PGE-2, COX-2 in tumor stroma. Also we have found tumor endothelial cells also express these inflammatory cytokines. Thus, we hypothesized that reactive oxygen species (ROS), which is produced in inflammation, may contribute to choromosomal abnormality or genomic instability in tumor endothelial cells (TECs). Green tea cathecin, EGCG has anti-oxidant and has anti-cancer effects. We have reported that EGCG inhibited VEGF-induced cell proliferation and migration in TEC, not in NEC (Cancer Sci. 2009). In this study, we addressed whether ROS in TEC is removed by EGCG, and analyzed the effects of ROS in TEC. By ROS accumulation, TEC could survive more compared to NEC. When ROS was induced by pyocyanin, VEGF/VEGFR signal was acitivated in TEC and it was inhibited by EGCG. In addition, ROS induced aneuploidy in NEC and EGCG inhibited VEGF and aneuploidy. These results suggested that ROS induced TEC abnormality via VEGF autocrine loop.
• 非小細胞肺癌幹細胞における分泌型miRNAの同定とその治療応用へ向けての解析

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  Date (from‐to) : 2012 -2012 

  Author : 加藤 達哉, 樋田 泰浩, 樋田 京子, 加賀 基知三, 松居 喜郎

   

  我々は、癌幹細胞に特化した「分泌型miRNA」に着目し、「非小細胞肺癌幹細胞における分泌型miRNAの同定とその治療応用へ向けての解析」というテーマで平成24年4月より研究を開始した。まず非小細胞肺癌における癌幹細胞の最も良いsorting方法を検討するため種々の方法を試みたが、中でも我々はsphere形成能を持つ癌細胞がマウス皮下腫瘍形成能を高率に有することに着目した。まず肺癌細胞株を用いてsphere形成能を持つ肺癌幹細胞の培養条件検討を行い、足場非依存性のためにpolyHEMA coatedを使用し、またserum-free、EGF+bFGF+ITS premixの培養液を用いて限界希釈を行ったのち培養を継続することで、同細胞群は約1ヶ月に渡りsphere形成能を維持したまま生育することを見出した。次に培養上清中の癌幹細胞における特異的な分泌型miRNAの検索として、マイクロアレイによる解析を行う方針とした。先に述べたsphere形成能を有する細胞群の培養上清と通常の肺癌細胞培養上清におけるmiRNA発現プロファイルを比較検討するため、両群の培養上清よりキットを用いてmiRNAの抽出を行った。細胞株レベルで条件検討が終了し、実際の臨床検体において同様の研究を開始する予定で倫理申請まで行っていたが、筆頭研究者である加藤達哉が海外へ留学することとなり、今回やむなく研究中止の申請をするに至った。
• Novel oncogenic mechanism of oral cancer mediated by HPV

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2011 -2012 

  Author : SATOH Akira, HIGASHINO Humihiro, KITAMURA Tetuya, HIDA Kyouko, TOTSUKA Yasunori

   

  In this study, we examined the export and stabilization of ARE-mRNA in oral cancer cells, which is caused by HPV. ARE-mRNA such as c-fos was stabilized in cells mediated by HPV oncogene products E6 and E7. The stabilization of ARE-mRNA in HPV positive cancer cells was down-regulated by HuR knockdown. These findings suggest that ARE-mRNA has some effect on the oncogenesis of cancer cells mediated by HPV.
• Targeting tumor metastasis by normalization of tumor blood vessels

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2011 -2012 

  Author : HIDA Kyoko, SHINDOH Masanobu, HIDA Yasuhiro, OHGA Noritaka

   

  We have isolated two different type of TECs (HM-TEC; isolated from highly metastatic tumor, and LM-TEC; isolated from low metastatic tumor) and found they are different in many aspects. Tumor cells were more attracted and adhesive to HM-TEC in vitro assay. They migrated through the HM-TEC monolayer most among all ECs. To address the question whether TECs contribute to tumor metastasis, low metastatic tumor cells were co-xenografted with each different type of EC (HM-TEC /LM-TEC/NEC) into nude mice. These results suggested that TECs in tumor microenvironment may activelyinduce tumor metastasis.
• Elucidation of invasive andmetastatic mechanism in oral cancer for application to molecular based diagnosisand gene therapy

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2009 -2012 

  Author : TAKATA Takashi, KUDO Yasusei, SINDOH Masanobu, HIDA Kyouko

   

  To elucidate invasive and metastatic mechanism in oral cancer for application to molecular based diagnosis and gene therapy, we conducted research in accordance with our projects. As a result, we identified Wnt5B and MMP10 as promoter of , microRNA-203 as suppressor of tumor invasion, Periostin as promoter of lymphangiogenesis, MMP-13 as promoter of angiogenesis. Additionally, we attempted to apply Periostin to Elisa for serum of oral cancer patients. Interestingly, we found upregulation of Periostin correlated with lymphonode metastasis status. Therefore, we showed Periostin could be a useful for molecular diagnostic marker in oral cancer.
• Basic research for oral cancer diagnosis estimated by HuR export

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2010 -2011 

  Author : TOTSUKA Yasunori, SHINDOH Masanobu, HIGASHINO Fumihiro, HIDA Kyouko, KITAMURA Tetsuya

   

  I this study, we examined the localization of HuR in oral cancer cells, the cytoplasmic expression of HuR and malignancy, the relation between cytoplasmic HuR and precancerous change. HuR and AU-rich containing mRNA were exported to the cytoplasm of oral cancer cells. Cytoplasmic HuR expression increased in the cancer which had high malignancy and in the precancerous change which showed poor diagnosis. These findings suggest that the localization of HuR is available for diagnostic tool of precancerous change.
• Mechanism of premetastatic niche formation and promotion of metastasis by cancer mesenchymal cells

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2009 -2011 

  Author : HIDA Yasuhiro, HIDA Kyoko, KONDO Satoshi

   

  Promotion of lung metastasis by interaction between tumor and tumor endothelial cells In this study, it was shown that tumor endothelial cells promote invasion and metastasis of tumor cells using cells labeled with fluorescein proteins. Tumor cells inoculated to mice with tumor endothelial cells circulated in peripheral blood and metastasized to lungs more than control conditions. The mechanism are enhanced migration of tumor cells towards tumor endothelial cells and transendothelial migration.
• Export of RNA-protein complex and oncogenesis

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2009 -2011 

  Author : ISHIKAWA Makoto, HIGASHINO Fumihiro, SHINNDOU Masanobu, HIDA Kyouko, KITAMURA Tetsuya

   

  In this study, we examined whether RNA binding proteins such as pp32 and HuR have oncogenic activity. AU-rich element(ARE) containing mRNA and HuR were exported to the cytoplasm of oral cancer cells. Although pp32 has potential to degrade HuR in the cytoplasm of cells, pp32r1 failed to do it. These results indicate that pp32r1 inhibits the degradation of HuR to stabilize ARE-mRNA and to transform cells
• Application of EPC for tissue regeneration

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2009 -2011 

  Author : ONO Mitsunobu, HIDA Kyoko, HIDA Yasuhiro, SHINDOH Masanobu, HIGASHINO Fumihiro, TOTSUKA Yasunori, KITAMURA Tetsuya

   

  EPC was isolated from peripheral blood using VEGFR2 and CD133 and they were cultured. Their angiogenic property was compared to normal endothelial cells(NEC). EPC showed higher levels of mRNA of growth factors and demonstrated activated angiogenic potential, ex, tube formation and cell migration in vitro. It was suggested that EPC may be contributed to tissue regeneration via their high angiogenic property.
• Exploitation of gene-targeting therapy based on the cellular and biological features of oral-carcinoma stem cell and tumor microenvironment.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2008 -2011 

  Author : TOTSUKA Yasunori, SHINDOH Masanobu, HIGASHINO Fumihiro, HIDA Kyoko, KITAMURA Tetsuya, OHBA Yusuke, MATSUMOTO Kenichi, OHIRO Yoichi

   

  We have shown that the tumor endothelial cells which exist in cancer stroma have a different gene background from normal endothelial cells. It has been becoming clear that epithelial-mesenchymal transition (EMT) plays an important role in tumor invasion and metastasis. It is expected that the investigation on the interaction of the oral squamous cell carcinoma cell in epithelium nature and a mesenchymal cell will contribute to the establishment of effective therapeutic method because stromal cells have the ability to supply nutrition and oxygen to a tumor. HuR protein usually exists in the nucleus, and HuR is participating to stabilize AU-rich element (ARE) mRNA. We showed that ARE mRNA is stabilized through HuR by a viral carcinogenesis system that concerns cell transformation. Oral environment has a unique features including distinct organs such as salivary glands. We identified that RANKL, an osteoclast inducer, expression was higher in oral environment. Furthermore, when oral cancer cells were implanted in oral region of nude mice, cancer cells proliferated intentionally, and EMT was observed. Tumor microenvironment is deeply correlated with cancer cell proliferation and invasion/metastasis. We have isolated endothelial cells in tumor stroma, and investigated the biological feature of tumor endothelial cells. As a result, tumor angiogenesis was suppressed by inhibiting Cox-2 known also for the factor inducing inflammation, and the resistance for chemothrapy was acquired by MDR1 upregulatin caused by VEGF signaling
• 血管新生阻害療法に対する腫瘍血管内皮細胞の抵抗性獲得の機序解析

  日本学術振興会：科学研究費助成事業 挑戦的萌芽研究

  Date (from‐to) : 2009 -2010 

  Author : 樋田 京子, 進藤 正信, 戸塚 靖則, 東野 史裕, 樋田 泰浩, 北村 哲也

   

  平成22年度はマウス腫瘍モデルを用いて,腫瘍血管新生阻害効果を解析した.同じ治療に対して抵抗性を獲得したマウスの腫瘍と感受性のあったマウスの腫瘍を用いてin situにおける血管内皮において前年度のin vitroの解析でピックアップされた血管内皮薬剤抵抗性関連因子の発現を解析した.さらに腫瘍微小環境が血管内皮細胞の薬剤抵抗性獲得に及ぼす影響に関して検討した. 腫瘍血管内皮細胞が薬剤抵抗性を獲得するメカニズムとして,腫瘍細胞と内皮細胞との相互作用による可能性,ならびに低酸素、薬剤による刺激などを考え,その機序について解析を行った.具体的にはがん細胞の培養上清による処理,またはがん細胞との共培養によって,血管内皮に薬剤抵抗性関連因子の発現亢進がおきるのかを検討した.がん細胞の培養上清によって正常血管内皮細胞の5-FUやpaclitaxelに対する薬剤抵抗性が表れること,さらに抵抗性関連遺伝子であるMDR-1の発現亢進などが起こること,MDR-1の転写因子であるYB-1の核内移行がおこることなどを見出した.さらに細胞内のAktのリン酸化亢進なども起こることをしめした.培養上清を熱処理するとこの現象は起こらなくなることから,培養上清中のサイトカインなどのタンパクが影響を及ぼす因子であることが示唆された.これらの結果より血管内皮細胞といえども環境により薬剤抵抗性を獲得しうるということが示唆された.
• Basic research for cancer therapy by knockdown of RNA binding protein

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2008 -2010 

  Author : HIGASHINO Fumihiro, SHINDOH Masanobu, TOTSUKA Yasunori, HIDA Kyouko, KITAMURA Tetsuya, KAKUGUCHI Wataru, KUROSHIMA Takeshi

   

  HuR is RNA binding protein which can protect ARE-mRNA from rapid degradation. In this study, we show that HuR-knockdown can change the malignant features of oral cancer cell. The findings show the potential of HuR-knockdown as an effective therapeutic approach.
• Development of novel antiangiogenic therapy based on tumor microenviroenment

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2008 -2010 

  Author : HIDA Kyoko, SHINDOH Masanobu, TOTSUKA Yasunori, HIGASHINO Fumihiro, HIDA Yasuhiro, KITAMURA Tetsuya

   

  To develope novel anti-angiogenic therapy reflecting on tumor microenvironment more, we compared gene expression between tumor endothelial cell (TEC) and normal endothelial cell (NEC) and identified TEC specific genes by DNA microarray analysis. We picked up these genes and analyzed the function of these molecules in TECs by knock down using siRNA. Knock down of some genes inhibited TEC survival and some inhibited cell migration on TEC. Furthermore, inhibition of several TEC markers showed tumor growth suppression in vivo. These results suggested that it enabled us to develope novel anti-angiogenic therapy targeting TEC specifically.
• Development of new screening system for anti-angiogenic drugs using tumor associated endothelial cells derived from renal cell carcinoma

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2008 -2010 

  Author : SHINOHARA Nobuo, HIDA Kyoko, NONOMURA Katsuya

   

  To develope new screening system for anti-angiogenic drugs, we isolated normal endothelial cell (NEC) and tumor endothelial cell (TEC) from human renal cell carcinoma xenografts and from dermis of normal mice, respectively. We analyzed the difference between TEC and NEC and investigate the potential of TEC for the screening system which reflect tumor microenvironment. Especially, we focused on several genes which were upregulated in TEC compared to NEC by DNA microarray analysis. We found these genes had strong association with abnormal phenotypes of TECs such as drug resistance, high viability, and migration. The specific phenotype in mouse TEC were also found in human TEC isolated from human renal cell carcinoma. These results enable us to approach new anti-angiogenic drug screening system and new anti-angiogenic therapy.
• Detection of cervical lymphnode micrometastasis in oral cancer

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2007 -2009 

  Author : ONO Mitsunobu, SHINDOH Masanobu, HIGASHINO Fumihiro, HIDA Kyoko, KITAMURA Tetsuya, OHIRO Yoichi, TOTSUKA Yasunori

   

  腫瘍の転移は悪性腫瘍の治療においてその予後に影響する最も重要な因子であり、口腔悪性腫瘍の大部分を占める扁平上皮がんの治療にあたっても、転移の有無が予後と深く関わっている。口腔扁平上皮がんの転移の大部分は所属リンパ節である頸部リンパ節へのリンパ行性転移で、その治療にあたっては原発巣とともに転移したリンパ節の制御が必須であり、とくに現在画像診断的に検出が困難である径5mm以下の微小リンパ節転移の有無を術前に知ることが可能であればその意義は大きい。今回の検索は、口腔がんにおける転移形質に関与する遺伝子/遺伝子産物の同定を行うことを第一の目標としている。 ヒト口腔正常粘膜上皮、白板症、口腔がん組織より試料を採取し、直ちに液体窒素で凍結保存を行い、以後の実験に用いた。試料の一部よりRNAを抽出しDNAマイクロアレイにより網羅的解析を行ったところ、数十の発現亢進する遺伝子および発現が低下した遺伝子が認められた。 これらの中で口腔がんで発現が亢進しているPim-1に注目して検索を行った。口腔扁平上皮がん細胞株6種全てにPim-1のタンパク発現が認められた。北海道大学病院で口腔扁平上皮がんと診断された早期がん症例39例についてPim-1の発現を検索したところ17例(44%)にPim-1発現が認められた。Pim-1発現症例は有意に所属リンパ節転移、遠隔臓器転移が多く、Pim-1発現と転移の関連性が示唆された。転移形質とPim-1の関連を探るため、siRNAを用いて検討を行った。その結果、Pim-1に対するsiRNA導入細胞では細胞運動能の低下がみられ、この際にRac1の活性が低下することが明らかになった。 以上の結果は、Pim-1はRac1の活性化により口腔がん細胞の運動能を高め、転移形質に深く関与しており、Pim-1をターゲットにした画像診断の有用性を示唆するものだった。
• 口腔がんの顎骨浸潤は抑制できるか?-EGFレセプターリン酸化阻害による効果-

  日本学術振興会：科学研究費助成事業 萌芽研究

  Date (from‐to) : 2007 -2008 

  Author : 戸塚 靖則, 進藤 正信, 大場 雄介, 北村 哲也, 樋田 京子, 東野 史裕, 津田 真寿美

   

  口腔がんの多くは舌や歯肉などの口腔内組織に発症するが、これらの臓器・組織は解剖学的に顎骨に近接しており、腫瘍の増大により容易に顎骨に浸潤する。この場合は、腫瘍の根治性の点から、顎骨離断術や部分切断術を施行せざるをえず、顔面の審美性や機能性に多大な障害を生じ、治癒後の社会生活やQOLに大きな影響を及ぼす。それ故、腫瘍の顎骨浸潤を抑制することが可能となれば、口腔がん患者にどって大きな福音となり、その開発が真に望まれている。 口腔癌細胞の多くは、epidemal growth factor (EGF) receptorを高頻度に発現しており、また顎下腺並びに唾液中には高濃度のEGFが存在していることが明らかになっている。従って、口腔癌の微小環境中に存在するEGFが、癌細胞に対してパラクライン的に作用し、骨浸潤に関与する形質発現に重要な影響を及ぼしていることが示唆される。 口腔がん細胞株HSC2, HSC3, HSC4, Ca9.22, SASを用いてEGFRの発現をWestern Blotで検討したところ、全てのがん細胞株でEGFRの発現亢進がみられた。これらの細胞をEGFで刺激した際、EGFRのリン酸化の亢進およびMAPKであるERK1/2, p38.JNKの活性化がみられた。ERK1/2はEGF依存的にPTHrPを発現亢進した。HSC2細胞にPTHrPのsiRNAを導入することで細胞増殖、浸潤活性の抑制がみられた。 このような所見は、口腔がん細胞の顎骨浸潤にはEGF/EGFRによるPTHrPの発現が重要な役割を演じていることを示唆するものだった。
• 口腔白板症の悪性化に関わる因子は何か?-HuRと細胞質移行タンパクの解析-

  日本学術振興会：科学研究費助成事業 萌芽研究

  Date (from‐to) : 2007 -2008 

  Author : 進藤 正信, 東野 史裕, 樋田 京子, 北村 哲也, 戸塚 靖則, 小林 正伸

   

  口腔の悪性腫瘍の大部分は扁平上皮がんが占めており、口腔がんの発症・進展は患者の咀嚼や発音などの機能障害だけでなく、審美障害を引き起こすため、その治療成績の改善は患者のQuolity of Life (QOL)の向上に密接な関係を有している。口腔は、可視化できる臓器であるため、これまで口腔扁平上皮がんの前がん病変として、所謂、白板症があることが知られている。 我々は、これまで口腔扁平上皮がんの発生・進展に関わっている遺伝子/遺伝子産物の研究を行ってきた。アデノウイルスのがん遺伝子産物を研究する過程で、E4orf6がAU-rich element (ARE)に結合することでmRNAを安定化させるHuRと結合し、核から細胞質へ移行することで細胞がん化を導くことを明らかにした。 口腔癌細胞株HSC3, Ca9.22を用いてHuRの発現をWestern blotおよび免疫染色で検索した。その結果、両細胞でHuRの発現は亢進し細胞質へ移行していることが明らかになった。HSC3の細胞増殖能はHuRの細胞質での発現がみられない歯肉線維芽細胞に比べ明らかに亢進していた。口腔がん細胞における駅EmRNAであるc-fos, c-myc mRNAの局在をin situ hybridization法で検討したところ正常細胞であるHGF(ヒト歯肉線維芽細胞)では核に局在していたが、口腔がん細胞では核および細胞質に存在し、ARE mRNAが核外輸送されていることが明らかになった。 さらに定量RT-PCR法でARE mRNAの安定化について検索した。その結果、口腔がん細胞ではc-fos, c-mycが細胞質内に蓄積・安定化されていた。 以上の結果は、口腔がん細胞ではHuRが核外へ輸送され、同時にARE mRNAも核外輸送・安定化されていることを示しており、HuRの細胞質への輸送局在を検討することにより、早期の細胞がん化を判定できる可能性が示された。
• RNA結合タンパクの局在と口腔がんの悪性度

  日本学術振興会：科学研究費助成事業 萌芽研究

  Date (from‐to) : 2007 -2008 

  Author : 東野 史裕, 進藤 正信, 戸塚 靖則, 樋田 京子

   

  AU(アデニン-ウラシル)-rich element (ARE)はc-fos、 c-myc、 COX-2などの細胞の増殖に関わる遺伝子のmRNAに存在し、AUUUAをコアとする繰り返し配列から成る領域である。我々はアデノウイルスのがん遺伝子産物E4orf6がARE-mRNAを核外輸送・安定化することにより細胞をがん化することをこれまでに見出している。E4orf6は発がんのみならず、がん細胞の悪性化に関わるタンパクでもありE4orf6によるARE-1nRNAや同時に輸送されるHuRの輸送は、細胞のがん化やがんの悪性化の指標となりうる。本研究の目的は口腔がん細胞でHuRタンパクの細胞質への局在と、そのがん細胞の悪性度との関連を調べ、HuRの局在が口腔がん細胞の悪性度の指標になるか検討することである。 口腔がん組織を用いて、HuRタンパクの局在を検討した。まず、HSC-3やCa9.22などの口腔がん培養細胞を用いてセルブロックを作成し、免疫染色の条件検討を行つた。次に、この染色条件で、手術材料から得られた切片を用いてHuRの免疫染色を行うによりHuRの局在を検討した。その結果、がん組織の細胞では細胞質側にもHuRタンパクが存在することがわかり、これに対して正常組織の細胞では主に核のみにHuRが局在していることが判明した。また、白板症の患者から得られたサンプルを用いて同様の検討を行つたところ、まだサンプル数は不十分であるが、予後不良のサンプルについてはHuRが核外輸送されていることも確認された。さらに、培養細胞を用いて、c-fos、 c-myc、 COX2などのARE-mRNAの局在をin situ hybridization法で検討しところ、がん細胞ではこれらのmRNAが核外輸送されていることも明らかになった。 以上の結果より、口腔がんでは、HuRやARE-mRNAが核外輸送されていることが明らかになり、HuRの局在が口腔がん細胞の悪性度の指標となりうることがわかった.
• Export and stabilization of AU-rich element containng mRNA in oral cancer cells

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2006 -2007 

  Author : HIGASHINO Fumihro, SHINDOH Masanobu, TOTSUKA Yasunori, KOBAYASHI Masanobu, HIDA Kyoko

   

  AU-rich element (ARE) exists in the mRNA transcribed from growth-related genes such as oncogene and cytokine. ARE-containning mRNA (ARE-mRNA) has been shown to be exported and stabilized in the cells transformed with virus oncoprotein. HuR is an RNA-binding protein which has ARE-mRNA are exported to the cytoplasm in oral cancer cells in a CRM1-independent manner and that the knockdown of HuR resulted in changing the characters of oral cancer cells. Immunohistochemical and biochemical analysis revealed that, in oral cancer cells such as HSC3 and Ca922, HuR existed in the cytoplasm and the nucleus whereas almost all HuR was located in the nucleus in normal cells. ARE-mRNAs were also exported and stabilized in oral cancer cells. The export of HuR and ARE-mRNA was not affected by Leptomycin B, which is the inhibitor of CRM1 dependent export pathway, in HSC3 cells. We developed a knockdown system, which is effective for oral cancer cells, using RNAi technique. In HuR-knockdown cells, the export of ARE-mRNA was inhibited. Additionally, those cells failed to make colonies in soft-agar. These findings suggest that HuR and ARE-mRNA are exported to the cytoplasm in oral carcinoma cells by purturbing the normal export pathway and that HuR knockdown has potential as a new therapeutic approach for oral cancer.
• Gene Targeting Therapy against tumor endothelial cells

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2006 -2007 

  Author : TOTSUKA Yasunori, SHINDOH Masanobu, HIDA Kyoko, HIGASHINO Fumihiro, KOBAYASHI Masanobu, OHIRO Yoichi

   

  Tumor is a genetic disorder, and gene targeting therapy for cancer cells has been conducted. However, tumor gene abnormalities arre complicated, and it is quite difficult to improve efficiency of cancer treatment targeting single gene product. On the other hand, since neovasculization in tumor stroma is essential for tumor survival and Growth, therapy for disturbing neovasculaizaion would be effective in cancer therapy. It has been considered that the blood vessels in tumor stroma was composed of normal endothelial cells; however, Hida et al. have reported that tumor endothelial cells have genetically varied character compared to the normal endothelial cells by isolation of tumor endothelial cells from mouth xenograft models of human cancers. We established human tumor endothelial cells by using magnetic beads method from the operation materials of oral carcinoma, renal carcinoma and lung carcinoma. These cells express CD34 and CD131, and confirmed as endothelial cells. RNA was extracted and DNA microarray has been carried out, and we have identified 70 genes that were upregulated in tumor endothelial cells. We confirmed the expression of these gene product by real-time RT-PCR, Western blotting and immunohistochemisty. Proliferation of tumor endothelial cells was decreased by introducing siRNA into the cells. In addition, we found that motility and proliferative activity of tumor endothelial cells were selectively decreased by Cox-2 inhibitor treatment. These results imply the significance of targeting tumor endothelial cells for favorable cancer therapy.
• Development of gene delivery system using nanocoloid and its application for gene targeting therapy

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2006 -2007 

  Author : SHINDOH Masanobu, KOBAYASHI Masanobu, FUGETSU Bunshi, HIGASHINO Fumihiro, HIDA Kyoko, KITAMURA Tetsuya

   

  Trials of gene therapies have been carried out for malignant tumors and congenital gene disorders,; however, there are problems such as the low efficiency and risk for applying human being, and ideal methods of gene therapy are not yet developed.. We designed nanocoloid that enclosed expression plasmid inside and, ligand peptide outside of small particle for development of a specific gene delivery system. When We searched expression of EGFR in oral squamous cell carcinoma cell lines (9.22, HSC2, HSC3, HSC4, Ca SAS) by Western blot, and all cell lines expressed EGFR We examined the binding affinity of EGFR and synthesized EGF peptide by BireCore X. As a result, it became clear that synthesized peptide bound to EGFR. Next, we examined the induction effect of nanocoloid to oral carcinoma cell lines. The efficiency of transfection was approximately 30%, and we aimed to improve the efficiency. We next examined the effects of adrenomedullin antagonist that has the ability to inhibit angiogenesis. The vector that has the sequence for inhibitory effects on aderenomedullin reduced the Growth of tumor xenografts in nude mice. These results imply that specific gene delivery for tumor environment could be the therapeutic agents for solid cancers.
• The significance of endothelial progenitor cell as a tumor marker in PBMC

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2006 -2007 

  Author : KAGA Kichizo, HIDA Yasuhiro, KONDO Satoshi, MIYAMOTO Masaki, HIDA Kyoko, SHICHINOHE Toshiaki

   

  Endothelial progenitor cell (EPC) is known as a significant material of angiogenesis in the tumor. EPC is released from the bone marrow and the number depends on the mediators secreted by tumor cells. Therefore, we hypothesized that EPC would show the status of tumor growth and indicates the prognosis of cancer patients. We constructed PEDF expressing lentivirus vector (Lv-PEDF) and established PEDF-overexpressiong cell lines HEC46-PEDF and TE8-PEDF. HEC46-PEDF and TE8-PEDF were implanted into nude mice subcutaneously and the growth was monitored After then, PBMC was extracted and the number of EPC was counted by using FACS. The number of EPC was increased in the mice with wild type HEC46 tumors or TE8 tumors, but not HEC46-PEDF and TE8-PEDF. Therefore, the data suggested that tumor cells secret factors inducing angiogenesis and they would be inhibited by PEDF Subsequently, we extracted PBMC from patients with tumor and assessed the number of PBMC and EPC.
• Analysis of Tumor-associated Endothelial Cell Biology

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2006 -2007 

  Author : HIDA Kyoko, SHINDOH Masanobu, TOTSUKA Yasunori, KOBAYASHI Masanobu, HIDA Yasuhiro, HIGASHINO Fumihiro

   

  Tumor angiogenesis is necessary for solid tumor progression and metastasis. Tumor blood vessels have been shown to differ from their normal counterparts, for example, by changes in morphology. An important concept in tumor angiogenesis is that tumor endothelial cells are assumed to be genetically normal, even though these endothelial cells are structurally and functionally abnormal. To date, many anti-angiogenic drugs have been developed, but, their therapeutic efficacy is not dramatical and it has been also reported to cause toxic side effects. To develop ideal antiangiogenic therapies, understanding tumor endothelial cell abnormalities is important. We have isolated tumor endothelial cells from mouse tumor xenografts and have shown that tumor endothelial cells are abnormal. Tumor endothelilal cells upregulate many genes, such as epidermal growth factor receptor (EGFR). Tumor endothelilal cells are also more sensitive to EGF. Furthermore, tumor endothelial cells were cytogenetically abnormal. Fluorescence in situ hybridization (FISH) analysis showed that freshly isolated uncultured tumor endothelial cells were aneuploid even when uncultured, contrary to current assumption. In marked contrast, normal skin endothelial cells were diploid and remained cytogenetically stable in culture. We conclude that tumor endothelial cells can acquire cytogenetic abnormalities while in the tumor microenvironment. Recently we analyzed the gene expression pattern of tumor endothelial cells and compared them to normal endothelial cells. We found the several genes which were upregulated in tumor endothelial cells. Further investigations are currently ongoing to investigate the significance of these tumor endothelial cell specific markers.
• study of tumor-associated endothelial cell specific markersbiology of tumor-associated endothelial cells biology of endothelial projenitor cells (EPC)

  Date (from‐to) : 2004
• 口腔がんにおける浸潤・転移の機構解明- 転写因子EIAFの機能分析-

  日本学術振興会：科学研究費助成事業 特別研究員奨励費

  Date (from‐to) : 1998 -1999 

  Author : 樋田 京子

Educational Activities

Teaching Experience

Social Contribution

Social Contribution

Social Contribution

• 血管と病気

  Date (from-to) : 2023/10/18-2023/10/18

  Role : Lecturer

  Event, Program, Title : アカデミックファンタジスタ, 市立札幌開成中等教育学校充実期（3－4年）課題研究スタートアップセミナー
• 「私のWork-Life Blend」

  Date (from-to) : 2021/03/01-2021/03/01

  Role : Lecturer

  Sponser, Organizer, Publisher  : 北海道大学病院男女共同参画推進室・北海道大学大学院歯学研究院FD委員会共催

  Event, Program, Title : 男女共同参画推進室講演会「キャリアを考える」
• 私のワークライフバランス－Try and Error－

  Date (from-to) : 2020/11/27-2020/11/27

  Role : Lecturer

  Sponser, Organizer, Publisher  : 九州大学

  Event, Program, Title : 令和2年度九州大学馬出地区4部局合同男女共同参画FD
• “継続は力なり～双子連れ留学，研究室開設の経験から”

  Date (from-to) : 2019/06

  Role : Lecturer

  Sponser, Organizer, Publisher  : 第57回日本小児歯科学会大会

  Event, Program, Title : 将来計画検討委員会・女性歯科医委員会合同企画シンポジウム
• “腫瘍血管の特性を活かした新しいがん治療法の開発を目指して”

  Date (from-to) : 2018/10/20

  Role : Lecturer

  Sponser, Organizer, Publisher  : これからのキャリアを語る医師と学生の会

  Event, Program, Title : これからのキャリアを語る医師と学生の会2018講演会「『がんと遺伝子』～私たちの研究が，患者さんの未来を変える」
• “研究指導者として、双子の親として”

  Date (from-to) : 2018/06/18

  Role : Lecturer

  Sponser, Organizer, Publisher  : 室蘭工業大学

  Event, Program, Title : キャリア形成のためのランチタイムセミナー第10回
• 「血管からがんを制す！」

  Date (from-to) : 2015/06-2015/06

  Role : Lecturer

  Sponser, Organizer, Publisher  : 北海道大学遺伝子病制御研究所

  Event, Program, Title : 北海道大学遺伝子病制御研究所一般公開サイエンストーク
• 「母親として，PI（Principle Investigator）として」

  Date (from-to) : 2012/07

  Role : Lecturer

  Sponser, Organizer, Publisher  : 広島大学男女共同参画推進室

  Event, Program, Title : 第9回広島大学女性研究者キャリアアップセミナー
• 講演「新しいがん治療法の開発に向けて」

  Date (from-to) : 2011/11

  Role : Lecturer

  Sponser, Organizer, Publisher  : 北海道歯科女医会

  Event, Program, Title : 平成23年度北海道歯科女医会総会
• 「プロジェクトリーダーとして,母親として」

  Date (from-to) : 2011/08

  Role : Lecturer

  Sponser, Organizer, Publisher  : 札幌南高等学校六華同窓会

  Event, Program, Title : 平成23年度第5回札幌南高等学校 六華ゼミ
• 「腫瘍血管を標的とする新しいがんの治療法開発を目指して」

  Date (from-to) : 2011/05

  Role : Lecturer

  Sponser, Organizer, Publisher  : 北海道大学歯学部同窓会北海道支部・札幌支部

  Event, Program, Title : 北海道大学歯学部同窓会北海道支部・札幌支部学術講演会
• 「カンペキでなくても、今できることを続けよう！」

  Date (from-to) : 2011/05

  Role : Appearance

  Sponser, Organizer, Publisher  : 北海道大学病院女性医師等支援事業

  Event, Program, Title : 北海道大学病院女性医師等支援室ホームページでの対談
• 北大女性研究者ショートリポート「家庭と研究の両立について」

  Date (from-to) : 2007/10

  Role : Lecturer

  Sponser, Organizer, Publisher  : 文部科学省科学技術振興調整費委託事業主催

  Event, Program, Title : 北海道大学女性研究者支援推進シンポジウム
• 理科実験の中学生への紹介ならびに口腔領域疾患について出前授業

  Date (from-to) : 2006/12/01

  Role : Organizing member

  Sponser, Organizer, Publisher  : 北大女性研究者支援室主催

  Event, Program, Title : 中高生理科系進路選択支援事業「理科してみよう！”Be Ambitious,中学生！」

Media Coverage

• 虫歯菌が血管に入るとがんの転移促す

  Date : 2022/10/24

  Program, newspaper magazine: NHKテレビ「ニュース（北海道）」

  Media report
• 洗口液に新型コロナ感染予防効果

  Date : 2022/09/01

  Program, newspaper magazine: 日本経済新聞（8月31日 電子版）

  Internet
• 市販の洗口液にコロナ予防効果

  Date : 2022/08/31

  Program, newspaper magazine: 北海道新聞

  Paper
• 科学新聞2022.7.1号“腫瘍血管の酸化LDL受容体 がん転移を促進”の記事掲載を紹介

  Date : 2022/08/04

  Publisher, broadcasting station: Science Japan by Japan Science and Technology Agency (JST)

  Internet
• 腫瘍血管の酸化LDL受容体 がん転移を促進

  Date : 2022/07/01

  Program, newspaper magazine: 科学新聞

  Paper
• 糖タンパク阻害で微小環境正常化

  Date : 2021/06

  Program, newspaper magazine: 北海道医療新聞

  Paper
• 北海道のがん研究最前線

  Date : 2020/10

  Writer: Myself

  Program, newspaper magazine: NHKテレビ「ほっとニュース北海道」

  Media report
• 血管異常が抗がん剤耐性原因

  Date : 2020/06

  Program, newspaper magazine: 北海道医療新聞

  Paper
• 抗癌剤による血管の薬剤耐性獲得メカニズムを解明～癌の薬剤耐性を克服する新たな治療法開発に期待～」

  Date : 2020/06

  Writer: Myself

  Publisher, broadcasting station: MedPeer

  Program, newspaper magazine: MEDICAL NEWS LINE

  Internet
• 第11回資生堂女性研究者サイエンスグラント 受賞

  Date : 2018/07

  Writer: Myself

  Program, newspaper magazine: 北海道新聞

  Paper
• 第49回 この人に聞く“生命に関わる仕事っておもしろいですか？”

  Date : 2017/10

  Publisher, broadcasting station: テルモ生命科学財団

  Program, newspaper magazine: 中高生と“いのちの不思議”を考える－生命科学DOKIDOKI研究室

  Internet
• 「第22回（2017年度）日本女性科学者の会 奨励賞」受賞

  Date : 2017/07

  Program, newspaper magazine: 朝日新聞

  Paper
• 「第22回（2017年度）日本女性科学者の会 奨励賞」受賞

  Date : 2017/07

  Program, newspaper magazine: 北海道医療新聞

  Paper
• がん転移 分泌タンパク質影響

  Date : 2016/07

  Program, newspaper magazine: 北海道新聞

  Paper
• がん転移の新しいメカニズムを解明

  Date : 2016/07

  Program, newspaper magazine: 北海道医療新聞

  Paper
• がん転移の仕組み解明

  Date : 2016/06/13

  Publisher, broadcasting station: NHKテレビ

  Program, newspaper magazine: 「ほっとニュース北海道」

  Media report
• がん転移の仕組み解明

  Date : 2016/06

  Publisher, broadcasting station: NHK

  Program, newspaper magazine: NHKテレビ「ほっとニュース北海道」

  Media report
• がん転移の仕組み解明

  Date : 2016/06

  Program, newspaper magazine: 日刊工業新聞

  Paper
• 血管を選択的に攻撃し，がんを兵糧攻めに

  Date : 2015/05

  Program, newspaper magazine: 北海道医療新聞

  Paper
• 新春展望 次世代の血管新生阻害療法の創生にむけて

  Date : 2015/01

  Program, newspaper magazine: 日経バイオテクONNLINE

  Internet
• 北海道大学遺伝子病制御研究所・金沢大学がん進展制御研究所ジョイントシンポジウム2014 での研究発表 新たな癌治療戦略を考えるには腫瘍血管内皮が正常血管内皮と異なることを理解すべき

  Date : 2014/11

  Program, newspaper magazine: 日経バイオテクONLINE

  Internet
• 「1分で知る豆医学」“血管 がん治療で注目”

  Date : 2013/11

  Program, newspaper magazine: 朝日新聞

  Paper
• “がんを制す！知られざる「血管の攻防戦」”

  Date : 2013/05

  Program, newspaper magazine: NHKテレビ「サイエンスZERO」

  Media report
• がん血管の特異性の解明と新しいがん治療・診断法への応用

  Date : 2013/01

  Publisher, broadcasting station: 日本学術振興会

  Program, newspaper magazine: 「科研費NEWSレター」（2012 VOL.3）

  Internet
• 北大，がんの転移能の違いで「腫瘍血管内皮細胞」にも違いがあることを解明

  Date : 2012/03

  Program, newspaper magazine: 科学新聞

  Paper
• 「北大，従来の常識を覆してがん内の血管も薬剤耐性が起こることを発見」

  Date : 2012/02

  Program, newspaper magazine: Yahooマイナビニュース

  Internet
• がんを養う血管においても薬剤耐性が起こることを発見

  Date : 2012/02

  Program, newspaper magazine: 科学新聞

  Paper
• 「北大，がんの転移能の違いで「腫瘍血管内皮細胞」にも違いがあることを解明」

  Date : 2012/02

  Program, newspaper magazine: Yahooマイナビニュース

  Internet
• 「知の達人たち」

  Date : 2011/09

  Program, newspaper magazine: 朝日新聞

  Paper
• 「北の知力」

  Date : 2010/06

  Publisher, broadcasting station: テレビ北海道

  Program, newspaper magazine: 北海道新聞ニュース

  Media report

Academic Contribution

• 第10回日本細胞外小胞学会学術集会『細胞外小胞の未来を見つめて』大会長

  Date (from-to) :2023/10/23-2023/10/24

  Role: Planning etc

  Type: Academic society etc

  Organizer, responsible person: 日本細胞外小胞学会
• 第65回歯科基礎医学会学術大会先端歯学シンポジウム「エキスパート研究の承継」オーガナイザー

  Date (from-to) :2023/09/17-2023/09/17

  Role: Planning etc

  Type: Academic society etc

  Organizer, responsible person: 歯科基礎医学会
• 第65回歯科基礎医学会学術大会アップデートシンポジウム「歯学基礎領域から発信する多角的アプローチからのがん研究最前線」オーガナイザー

  Date (from-to) :2023/09/17-2023/09/17

  Role: Panel chair etc

  Organizer, responsible person: 歯科基礎医学会
• 第81回日本癌学会総会 シンポジウム「がん微小環境：転移と浸潤」オーガナイザー

  Date (from-to) :2022/09/30-2022/09/30

  Role: Planning etc

  Type: Academic society etc

  Organizer, responsible person: 日本癌学会
• 第64回歯科基礎医学会学術大会 日本学術会議シンポジウム（市民公開）Science Council of Japan Symposium「口腔と全身のネットワーク～脈管系から生命現象を理解する」, オーガナイザー

  Date (from-to) :2022/09/17-2022/09/17

  Role: Planning etc

  Type: Academic society etc

  Organizer, responsible person: 歯科基礎医学会
• 第121回北海道癌談話会 春季シンポジウム

  Date (from-to) :2021/06/26

  Role: Planning etc

  Type: Competition etc

  Organizer, responsible person: 北海道癌談話会
• 第53回北海道病理談話会

  Date (from-to) :2020/10/10

  Role: Planning etc

  Type: Academic society etc

  Organizer, responsible person: 北海道病理談話会
• 第58回歯科基礎医学会学術大会「サテライトシンポジウム10 “機能と形態の統合的アプローチによる血管・唾液腺・骨の共創的研究”

  Date (from-to) :2016/08/24

  Role: Planning etc

  Type: Competition etc

  Organizer, responsible person: 歯科基礎医学会
• 公開シンポジウム「血管を標的とするナノ医療の実用化に向けた拠点形成～がんを始めとする国民病を血管から治療する～」

  Date (from-to) :2014/06/27

  Role: Planning etc

  Type: Competition etc

  Organizer, responsible person: 血管を標的とするナノ医療の実用化に向けた拠点
• 公開シンポジウム「血管を標的とする革新的医薬分子送達法の基盤技術の確立」

  Date (from-to) :2012/01/12

  Role: Planning etc

  Type: Competition etc

  Organizer, responsible person: 北大部局横断型連携プロジェクト「血管を標的とする革新的医薬分子送達法の基盤技術の確立」
• 第64回歯科基礎医学会学術大会 アップデートシンポジウム「歯科基礎領域におけるがん研究フロンティア」 オーガナイザー 2022.9.18（徳島）台風のため誌上開催

  Role: Planning etc

  Type: Academic society etc

  Organizer, responsible person: 歯科基礎医学会