研究者データベース
| 石原 すみれ(イシハラ スミレ) |
| 先端生命科学研究院 生命機能科学研究部門 分子細胞生物学分野 |
| 助教 |
Last Updated :2023/12/05
研究活動情報
論文
- Koya Yoshizawa, Akira Matsura, Masaya Shimada, Sumire Ishida‐Ishihara, Fuyu Sato, Takahiro Yamamoto, Kan Yaguchi, Eiji Kawamoto, Taruho Kuroda, Kazuya Matsuo, Nobuyuki Tamaoki, Ryuichi Sakai, Yasuhito Shimada, Mithilesh Mishra, Ryota UeharaMolecular Oncology 17 6 1148 - 1166 2023年02月11日 [査読有り]
Tetraploidy is a hallmark of cancer cells, and tetraploidy‐selective cell growth suppression is a potential strategy for targeted cancer therapy. However, how tetraploid cells differ from normal diploids in their sensitivity to anti‐proliferative treatments remains largely unknown. In this study, we found that tetraploid cells are significantly more susceptible to inhibitors of a mitotic kinesin (CENP‐E) than are diploids. Treatment with a CENP‐E inhibitor preferentially diminished the tetraploid cell population in a diploid–tetraploid co‐culture at optimum conditions. Live imaging revealed that a tetraploidy‐linked increase in unsolvable chromosome misalignment caused substantially longer mitotic delay in tetraploids than in diploids upon moderate CENP‐E inhibition. This time gap of mitotic arrest resulted in cohesion fatigue and subsequent cell death, specifically in tetraploids, leading to tetraploidy‐selective cell growth suppression. In contrast, the microtubule‐stabilizing compound paclitaxel caused tetraploidy‐selective suppression through the aggravation of spindle multipolarization. We also found that treatment with a CENP‐E inhibitor had superior generality to paclitaxel in its tetraploidy selectivity across a broader spectrum of cell lines. Our results highlight the unique properties of CENP‐E inhibitors in tetraploidy‐selective suppression and their potential use in the development of tetraploidy‐targeting interventions in cancer. - Sumire Ishida-Ishihara, Ryota Takada, Kazuya Furusawa, Seiichiro Ishihara, Hisashi HagaScientific reports 12 1 20269 - 20269 2022年11月24日 [査読有り]
Cell-containing collagen gels are one of the materials employed in tissue engineering and drug testing. A collagen gel is a useful three-dimensional (3D) scaffold that improves various cell functions compared to traditional two-dimensional plastic substrates. However, owing to poor nutrient availability, cells are not viable in thick collagen gels. Perfusion is an effective method for supplying nutrients to the gel. In this study, we maintained hepatocytes embedded in a 3D collagen gel using a simple pump-free perfusion cell culture system with ordinary cell culture products. Flow was generated by the difference in water level in the culture medium. Hepatocytes were found to be viable in a collagen gel of thickness 3.26 (± 0.16 S.E.)-mm for 3 days. In addition, hepatocytes had improved proliferation and gene expression related to liver function in a 3D collagen gel compared to a 2D culture dish. These findings indicate that our perfusion method is useful for investigating the cellular functions of 3D hydrogels. - S. Ishida-Ishihara, M. Akiyama, K. Furusawa, I. Naguro, H. Ryuno, T. Sushida, S. Ishihara, H. HagaJournal of Cell Science 133 14 2020年 [査読有り]
One of the fundamental processes in morphogenesis is dome formation, but many of the mechanisms involved are unexplored. Previous in vitro studies showed that an osmotic gradient is the driving factor of dome formation. However, these investigations were performed without extracellular matrix (ECM), which provides structural support to morphogenesis. With the use of ECM, we observed that basal hypertonic stress induced stable domes in vitro that have not been seen in previous studies. These domes developed as a result of ECM swelling via aquaporin water transport activity. Based on computer simulation, uneven swelling, with a positive feedback between cell stretching and enhanced water transport, was a cause of dome formation. These results indicate that osmotic gradients induce dome morphogenesis via both enhanced water transport activity and subsequent ECM swelling. - Y. Kumagai, J. Nio-Kobayashi, S. Ishida-Ishihara, H. Tachibana, R. Omori, A. Enomoto, S. Ishihara, H. HagaBiochemical and Biophysical Research Communications 514 4 1115 - 1121 2019年 [査読有り]
Cancer cells can invade as a population in various cancer tissues. This phenomenon is called collective invasion, which is associated with the metastatic potential and prognosis of cancer patients. The collectiveness of cancer cells is necessary for collective invasion. However, the mechanism underlying the generation of collectiveness by cancer cells is not well known. In this study, the phenomenon of contact following, where neighboring cells move in the same direction via intercellular adhesion, was investigated. An experimental system was created to observe the two-dimensional invasion using a collagen gel overlay to study contact following in collective invasion. The role of integrin-β1, one of the major extracellular matrix (ECM) receptors, in contact following was examined through the experimental system. Integrin-β1 was localized to the intercellular site in squamous carcinoma cells. Moreover, the intercellular adhesion and contact following were suppressed by treatment of an integrin-β1 inhibitory antibody. ECM proteins such as laminin-332 and type-XVII collagen were also localized to the intercellular site and critical for contact following. Collectively, it was demonstrated that the activity of integrin-β1 and expression of ECM proteins in the intercellular site promote contact following in the collective invasion of a cancer cell population. - X. Wang, A. Enomoto, L.Weng, Y. Mizutani, S. Abudureyimu, N. Esaki, Y. Tsuyuki, C. Chen, S. Mii, N. Asai, H. Haga, S. Ishida, K. Yokota, M. Akiyama, M. TakahashiCancer Science 109 11 3643 - 3656 2018年 [査読有り]
Pathological observations show that cancer cells frequently invade the surrounding stroma in collective groups rather than through single cell migration. Here, we studied the role of the actin-binding protein Girdin, a specific regulator of collective migration of neuroblasts in the brain, in collective cancer cell migration. We found that Girdin was essential for the collective migration of the skin cancer cell line A431 on collagen gels as well as their fibroblast-led collective invasion in an organotypic culture model. We provide evidence that Girdin binds to β-catenin that plays important roles in the Wnt signaling pathway and in E-cadherin-mediated cell-cell adhesion. Girdin-depleted cells displayed scattering and impaired E-cadherin-specific cell-cell adhesion. Importantly, Girdin depletion led to impaired cytoskeletal association of the β-catenin complex, which was accompanied by changes in the supracellular actin cytoskeletal organization of cancer cell cohorts on collagen gels. Although the underlying mechanism is unclear, this observation is consistent with the established role of the actin cytoskeletal system and cell-cell adhesion in the collective behavior of cells. Finally, we showed the correlation of the expression of Girdin with that of the components of the E-cadherin complex and the differentiation of human skin cancer. Collectively, our results suggest that Girdin is an important modulator of the collective behavior of cancer cells. - Mathematical model of collective cell migrations based on cell polarity.M. Akiyama, T. Sushida, S. Ishida, H. HagaDevelopment, Growth & Differentiation 59 5 471 - 490 2017年 [査読有り]
- TRIM27/MRTF-B-dependent integrin β1 expression defines leading cells in cancer cell collectives.T. Kato, A. Enomoto, T. Watanabe, H. Haga, S. Ishida, Y. Kondo, K. Furukawa, S. Mii, L. Weng, M. Ishida-Takagishi, M. Asai, N. Asai, K. Kaibuchi, Y. Murakumo, M. TakahashiCell Reports 7 1 - 12 2014年 [査読有り]
- S. Ishida, R. Tanaka, N. Yamaguchi, G. Ogata, T. Mizutani, K. Kawabata, H. HagaPLoS ONE 9 8 e99655 Public Library of Science 2014年 [査読有り]
Lumen formation is important for morphogenesis; however, an unanswered question is whether it involves the collective migration of epithelial cells. Here, using a collagen gel overlay culture method, we show that Madin-Darby canine kidney cells migrated collectively and formed a luminal structure in a collagen gel. Immediately after the collagen gel overlay, an epithelial sheet folded from the periphery, migrated inwardly, and formed a luminal structure. The inhibition of integrin-beta 1 or Rac1 activity decreased the migration rate of the peripheral cells after the sheets folded. Moreover, lumen formation was perturbed by disruption of apical-basolateral polarity induced by transforming growth factor-beta 1. These results indicate that cell migration and cell polarity play an important role in folding. To further explore epithelial sheet folding, we developed a computer-simulated mechanical model based on the rigidity of the extracellular matrix. It indicated a soft substrate is required for the folding movement.
受賞
共同研究・競争的資金等の研究課題
- 「液体」上で細胞を飼う：器官構築の根幹に迫る新規アプローチ日本学術振興会：科学研究費助成事業 特別研究員奨励費研究期間 : 2017年04月 -2019年03月代表者 : 石原(石田)すみれ, 石田 すみれ動物の複雑な体構造は，上皮組織の座屈や折りたたみ・伸長といった単純な変形が組み合わされることにより形成される．ドーム形成は上皮組織の折りたたみのひとつであるが，ドーム形成を引き起こす要因はほとんど分かっていない．先行研究により浸透圧勾配が要因として提唱されているものの，その直接的証拠は報告されていない．そこで我々は浸透圧勾配によってドーム形成が誘引されるかとその仕組みとをインビトロで調べた． 本研究では，液体培養基質であるマトリゲルをゲニピンという架橋剤で処理して使用した．このゲニピン処理マトリゲル（以下，GPM）上の上皮細胞シートへ浸透圧勾配を与えると，細胞シートは内部がGPMで満たされたドームへと変形した． 次に，浸透圧勾配がGPM上でドーム形成を起こす仕組みを調べた．3Dライブイメージングの結果，ドーム形成時にはGPMが膨潤していることがわかった．ゲルは内部の溶液の塩濃度によって膨潤度が変わる．また，細胞の水チャネル・アクアポリン（AQP）は水を高張側に輸送する．そのため，ドーム形成時もAQPによって水がGPMへと輸送されてGPM中の塩濃度が下がり、膨潤が起こっている可能性がある．この可能性を検証するため，AQP阻害剤を投与した．その結果，膨潤が抑制されドームもできなかった． さらに我々は，ドーム構造の力学的機序を調べた．ミオシンはリン酸化により細胞内張力を発生させる．このミオシンの阻害剤（ブレビスタチン, Y27632）を投与したところ，ドーム形成が阻害された．現在は，細胞内張力の局在を調べるため，リン酸化ミオシンの免疫蛍光染色に取り組んでいる． これらの事実は，浸透圧勾配によってGPM上の細胞シートにドーム形成が誘引されることを示した重要な結果である．今後は，細胞内張力の局在明らかにし，来年度中に成果を論文にまとめる予定である．
- 科学の芽を育む実験教室北海道大学：北大元気プロジェクト研究期間 : 2011年 -2012年