ヒト癌浸潤におけるアダプター分子クラックの解析

• Project/Area Number: 18013003
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Hokkaido University
• Principal Investigator: 田中 伸哉 Hokkaido University, 大学院・医学研究科, 准教授 (70261287)
• Co-Investigator(Kenkyū-buntansha): 西原 広史 北海道大学, 大学院・医学研究科, 助教 (50322805)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥11,000,000 (Direct Cost: ¥11,000,000); Fiscal Year 2007: ¥5,300,000 (Direct Cost: ¥5,300,000); Fiscal Year 2006: ¥5,700,000 (Direct Cost: ¥5,700,000)
• Keywords: crk / シグナル伝達 / アダプター / 構造解析 / NMR / CRK / 細胞骨格 / G蛋白質

Research Abstract

本研究ではアダプター分子Crkの癌化における役割を解析し、癌腫、肉腫、脳腫瘍など様々なヒト癌細胞株を用いてsiRNA法にてCrk knockdown細胞株を樹立した。何れの種類のCrk knockdown細胞においても、接着能、浸潤能、足場非依存性増殖能、in vivo造腫瘍能など、癌細胞の悪性化を示す指標に、著明な減少がみられ、Crkはヒト卵巣癌、軟部肉腫、脳腫瘍において、悪性化に必須の分子であることが明かとなった(Oncogene,25, 2006:M01. Cancer Res.,7, 2006)。また、Crkの癌化におけるシグナル伝達メカニズムを詳細に解析するために、NMRを用いた構造解析を行ったところ、Crk-Iはflexibleな分子で恒常的にシグナル標的分子と結合しているが、Crk-IIは、Grb2などのアダプタ一分子とは異なり、3つのSH領域がコンパクトな構造をとっており、この構造が2つのSH3領域の間にある15アミノ酸によって制御されることが判明し、この領域をISC (inter SH3 core)と命名した。ISCの変異体は、標的分子への親和性が回復し、また、細胞増殖能もCrk-Iと同様のレベルまで増加した。したがって、Crk-IIのcompactなstructureがシグナルの制御に関与することが判明した(Nature Struct. Mol. Biol.,2007, (In press)。今後は本研究で明らかにされた構造に基づきCrkのシグナル伝達を抑制する薬剤開発を目指す。

Research Products

• [Journal Article] Structural basis for the transforming activity of human cancer-related signaling adaptor protein CRK. (2007)
  • Author(s): Kobashigawa, Y., Sakai, M., Naito, M., Yokochi, M., Kumeta, H., Makino, Y., Ogura, K., *Tanaka, S., *Inagaki, F.(*co-corresponding author)
  • Journal Title: Nature Strct&Mol.Biol. 6 Pages: 503-510
  • NAID: 40015830831
  • Peer Reviewed
• [Journal Article] O^6-methylguanine-DNA methyltransferase is downregulated in transformed astrocyte cells:implications for anti-glioma therapies. (2007)
  • Author(s): Sasai, K., Akagi, T., Aoyanagi, E., Tabu, K., Kaneko, S., Tanaka, S.
  • Journal Title: Mol.Cancer 6 Pages: 36-36
  • Peer Reviewed
• [Journal Article] Expression of OLIG2 suppresses the motility of human glial tumor cell line via RhoA activation (2007)
  • Author(s): Tabu, K., Ohba, Y., Suzuki, T., Makino, Y., Ohnishi, A., Sakai, M., Watanabe, T., Kimura, T., Tanaka, S., Sawa, H.
  • Journal Title: Mol.Cancer Res (In press)
  • Peer Reviewed
• [Journal Article] Enteritis necroticans'pigbel'in a Japanese diabetic adult (2007)
  • Author(s): Matsuda, T., Okada, Y., Inagi, E., Tanabe, Y., Shimizu, Y., Nagashima, K., Sakurai, J., Nagahama, M., Tanaka, S.
  • Journal Title: Pathol.Inter 57 Pages: 622-626
  • Peer Reviewed
• [Journal Article] Primary Gastric Hodgkin's Lymphoma Expressing a B-Cell Profile Including Oct-2 and Bob-1 Proteins. (2007)
  • Author(s): Saito, M., Tanaka, S., Mori, A., Toyoshima, N., Irie, T., Morioka, M.
  • Journal Title: Int J Hematol 85 Pages: 421-425
  • Peer Reviewed
• [Journal Article] Type I interferon prolongs cell cycle progression via p21 WAF1/CIP1 induction in human colon cancer cells. (2007)
  • Author(s): Katayama, T., Nakanishi, K., Nishihara, H., Kamiyama, N., Nakagawa, T., Kamiyama, T., Iseki, K., Tanaka, S., Todo, S.
  • Journal Title: International Journal of oncology 31 Pages: 613-620
  • Peer Reviewed
• [Journal Article] Signaling adaptor protein Crk is indispensable for malignant feature of gliblastoma cell line KMG4. (2007)
  • Author(s): Wang, L., Tabu, K., Kimura, T., Tsuda, M., Linghu, H., Tanino, M., Kaneko, S., Nishihara, H., Tanaka, S.
  • Journal Title: Biochem Biophys Res Commun 362 Pages: 976-981
  • Peer Reviewed
• [Journal Article] Immunohistochemical and gene rearrangement studies of central nervous system lymphomatoid granulomatosis. (2007)
  • Author(s): Nishihara, H., Tateishi, U., Itoh, T., Nagashima, K., Tanaka, S.
  • Journal Title: Neuropathology 27 Pages: 413-418
  • NAID: 10021236336
  • Peer Reviewed
• [Journal Article] Elmol inhibits the ubiquitylation of Dock180 recruited onto plasma membrance. (2006)
  • Author(s): Makino, Y., Tsuda, M., Ichihara, S., Watanabe, T., Sakai, M., Sawa, H., Nagashima, K., Hatakeyama, S., Tanaka, S.
  • Journal Title: J. Cell Sci. 119 Pages: 923-932
• [Journal Article] A novel function of OLIG2 to suppress human glial tumor cell growth via p27Kip1 transactivation. (2006)
  • Author(s): Tabu, K., Ohnishi, A., Sunden, Y., Suzuki, T., Tsuda, M., Tanaka, S., Sakai, M., Nagashima, K., Sawa.H.
  • Journal Title: J, Cell Sci. 119 Pages: 1433-1441
• [Journal Article] IGF2 is critical for tumorigenesis by synovial sarcoma oncoprotein SYT-SSX1. (2006)
  • Author(s): Sun, Y., Gao, D., Liu, Y., Huang, J., Lessnick, S., Tanaka, S.
  • Journal Title: Oncogene 25 Pages: 1042-1052
• [Journal Article] Azathioprine suppresses ERM-dependent T cell-APC conjugation through inhibition of Vav guanosine exchange activity on Rac proteins. (2006)
  • Author(s): Poppe, D., Tiede, I., Fritz, G., Becker, C., Bartsch, B., Wirtz, S., Strand, D., Tanaka, S., Galle, PR., Bustelo, XR., Neurath, MF.
  • Journal Title: J. Immunol. 176 Pages: 640-651
• [Journal Article] Characterization and application of polyclonal antibodies that specifically recognize JC virus large T antigen. (2006)
  • Author(s): Sunden, Y., Suzuki, T., Orba, Y., Umemura, T., Asamoto, M., Nagashima, K., Tanaka, S., Sawa, H.
  • Journal Title: Acta Neuropathol. (Berl) 111 Pages: 379-387
• [Journal Article] Involvement of adaptor protein Crk in malignant feature of human ovarian cancer cell line MCAS. (2006)
  • Author(s): Linghu, H., Tsuda, M., Makino, Y., Sakai, M., Watanabe, T., Ichihara, S., Sawa, H., Nagashima, K., Mochizuki, N., Tanaka, S.
  • Journal Title: Oncogene 25 Pages: 3547-3556
• [Journal Article] Adaptor molecule Crk is required for sustained phosphorylation of Gabl and HGF-induced cell motility of human synovial sarcoma cell lines (2006)
  • Author(s): Wanatabe, T., Tsuda, M., Makino, Y., Ichihara, S., Sawa, H., Mochizuki, N., Nagashima, K., Tanaka, S.
  • Journal Title: Mol. Cancer Res. 7 Pages: 499-510