細胞接着シグナルを介したがん細胞の浸潤制御機構

• Project/Area Number: 12215157
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka Bioscience Institute
• Principal Investigator: 佐邊 壽孝 (財)大阪バイオサイエンス研究所, 分子生物学部門, 研究部長 (40187282)
• Co-Investigator(Kenkyū-buntansha): 永渕 昭良 熊本大学, 発生医学研究センター・初期発生分野, 教授 (80218023) ; 広橋 説雄 国立がんセンター, 研究所, 所長 (70129625)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥100,000,000 (Direct Cost: ¥100,000,000); Fiscal Year 2004: ¥22,000,000 (Direct Cost: ¥22,000,000); Fiscal Year 2003: ¥22,000,000 (Direct Cost: ¥22,000,000); Fiscal Year 2002: ¥24,000,000 (Direct Cost: ¥24,000,000); Fiscal Year 2001: ¥14,000,000 (Direct Cost: ¥14,000,000); Fiscal Year 2000: ¥18,000,000 (Direct Cost: ¥18,000,000)
• Keywords: インテグリン / カドヘリン / 細胞運動 / 浸潤 / 乳癌 / カテニン / P120 / F9 / 浸潤性 / Rho-family GTPases / パキシリン / βカテニン / プラコグロビン / paxillin / RhoA / RhoGAP / membrane ruffle / アクチン細胞骨格再構成 / チロシンリン酸化 / p130Cas / ARFGAP / c-Src

Research Abstract

がん細胞、特に上皮由来癌の浸潤機序とそれに関わる分子機構を明らかにするため、インテグリン裏打ち蛋白質とカドヘリン裏打ち蛋白質に着目し解析を進めている。これまでの研究を踏まえ本年度は、パキシリンとその結合性ArfGAPを含む、乳癌の浸潤活性に重要な蛋白質複合体を同定し、その存在が乳癌(細胞株、病理切片)の浸潤形質と良く一致すること、siRNA法による個々の蛋白質発現の阻害や蛋白質複合体の形成をペプチド等によって阻害することによって乳癌の浸潤と転移を著しく阻害できることを示し、この蛋白質複合体が抗癌剤開発の分子標的としての評価対象になり得ることを提示した(佐邊)。一方、懸案であるEカドヘリンとNカドヘリンとの動態の差異がどのようにして癌の浸潤形質獲得と連関するのかに関して、カドヘリンの動態の差異と癌の浸潤活性の両方にArf6が関与することを明らかにし、細胞間接着形成においてArf6の活性制御に関わる細胞表面受容体も同定した。分子機序に関しては現在解析中である(佐邊)。
カドヘリン裏打ち蛋白質に関しては、F9細胞を用いて細胞の接着形態における役割を検討し、β-カテニンとプラコグロビン両者を欠損させると細胞接着能が減弱することを示した(永渕)。

Research Products

• [Journal Article] Expression of AMAP1, an ArfGAP, provides novel targets to inhibit breast cancer invasive activities. (2005)
  • Author(s): Y.Onodera, S.Hashimoto, A.Hashimoto, M.Morishige, Y.Mazaki, A.Yamada, E.Ogawa, M.Adachi, T.Sakurai, T.Manabe, H.Wada, N.Matsuura, H.Sabe
  • Journal Title: EMBO J. (in press)
• [Journal Article] A novel mode of action of an ArfGAP, PAG3/AMAP2/Papα, in Arf6 function. (2004)
  • Author(s): S.Hashimoto, A.Hashimoto, A.Yamada, C.Kojima, H.Yamamoto, T.Tsutsumi, M.Higashi, A.Mizoguchi, R.Yagi, H.Sabe
  • Journal Title: J.Biol.Chem. 279・36 Pages: 37677-37684
• [Journal Article] Phosphorylation of paxillin tyrosines 31 and 118 controls polarization and motility of lymphoid cells and is PMA-sensitive. (2004)
  • Author(s): L.Y.Romanova, S.hashimoto, K.-O.Chay, M.V.Blagosklonny, H.Sabe, J.F.Mushinski.
  • Journal Title: J.Cell Sci. 117・17 Pages: 3759-3768
• [Journal Article] Requirement for Arf6 in breast cancer invasive activities. (2004)
  • Author(s): S.Hashimoto, Y.Onodera, A.Hashimoto, M.Tanaka, N.Hamaguchi, A.Yamada, H.Sabe
  • Journal Title: Proc.Natl.Acad.Sci.USA 101・17 Pages: 6647-6652
• [Journal Article] A role played by a subset of integrin-signaling molecules in cadherin-based cell-cell adhesions. (2004)
  • Author(s): H.Yano, Y.Mazaki, K.Kurokawa, S.K.Hanks, M.Matsuda, H.Sabe
  • Journal Title: J.Cell Biol. 166・2 Pages: 283-295
• [Journal Article] Regulation of Bin1 SH3 domain binding by phosphoinositides. (2004)
  • Author(s): C.Kojima, A.Hashimoto, I.Yabuta, M.Hirose, S.Hashimoto, Y.Kanaho, H.Sumimoto, T.Ikegami, H.Sabe
  • Journal Title: EMBO J. 23・22 Pages: 4413-4422
• [Patent(Industrial Property Rights)] ヒト癌細胞の運動と浸潤を阻害する物質およびそのスクリーニング方法 (2004)
  • Inventor(s): 佐邊 壽孝, 橋本 茂
  • Industrial Property Rights Holder: (財)大阪バイオサイエンス研究所
  • Filing Date: 2004-09-06
• [Publications] H.Watanabe, T.Shimizu, J.Nishihira, R.Abe, T.Nakayama, M.Taniguchi, H.Sabe, T.Ishibashi, H.Shimizu.: "Ultraviolet A-induced production of matrix metalloproteinase-1 is mediated by macrophagemigration inhibitory factor in human dermal fibroblasts."J Biol.Chem.. 279・3. 1676-1683 (2004)
• [Publications] H.Sabe.: "A requirement for Arf6 in cell adhesion and migration, and cancer cell invasion."J.Biochem.Minireview. 134・4. 485-489 (2003)
• [Publications] C.Didier, L.Broday, A.Bhoumik, S.Israeli, S.Takahashi, K.Nakayama, S.M.Thomas, C.E.Turner, S.Henderson, H.Sabe, Z.Ronai.: "RNF5-a RING finger protein that regulates cell motility by targeting paxillin ubiquitination and altered localization."Mol.Cell Biol.. 23・15. 5331-5345 (2003)
• [Publications] H.Sabe.: "The ARF book"(Ed.R.Kahn)Kluwer Academic Publishers.(in press).
• [Publications] A.Tsubouchi, J.Sakakura, R.Yagi, Y.Mazaki, E.Schaefer, H.Yarno, H.Sabe: "Tyrosine phosphorylation of paxillin is important for microtubule organizing center positioning and directional persistency in motile epithelial cells"J.Cell Biol.. 159・4. 673-683 (2002)
• [Publications] Y.Yamamoto, T.Maruyama, N.Sakai, R.Sakurai, A.Shimizu, T.Hamatani, H.Masuda, H.Uchida, H.Sabe, Y.Yoshimura: "Expression and subcellular distribution of the active form of c-Src tyrosine kinase in differentiating human endometrial stromal cells"Mol.Hum.Reprod.. 8・12. 1117-1124 (2002)
• [Publications] T.Oshiro, S.Koyama, S.Sugiyama, A.Kondo, Y.Onodera, T.Asahara, H.Sabe, A.Kikuchi: "Interaction of POB1, a downstream molecule of small G protein Ral, with PAG2, a paxillin binding protein, is involved in cell migration"J.Biol.Chem.. 277・41. 38618-38626 (2002)
• [Publications] A.J.Woods, M.S.Roberts, J.Choudhary, S.T.Barry, Y.Mazaki, H.Sabe, S.J.Morley, D.R.Critchley, J.C.Norman: "Paxillin associates with poly(A)-binding protein lat the dense ER and the leading edge of migrating cells"J.Biol.Chem.. 277・8. 6428-6437 (2002)
• [Publications] T.Iwasaki, A.Nakata, M.Mukai, K.Shinkai, H.Yano, H.Sabe, E.Schaefer, M.Tatsuta, T.Tsujimura, N.Terada, E.Kakishita, H.Akedo: "Involvement of phosphorylation of Tyr-31 and Tyr-118 of paxillin in MM1 cancer cell migration"Int.J.Cancer. 97・3. 330-335 (2002)
• [Publications] H.Sabe: "In 'The ARF book'(Ed. R.Kahn)"Kluwer Academic Publishers (in press). 18
• [Publications] Hashimoto, S., Tsubouchi, A., Mazaki, Y., Sabe, H.: "Interaction of paxillin with p21-activated kinase (PAK) Association of paxillin alpha with the kinase-inactive and the Cdc42-activated forms of PAK3"J.Biol.Chem.. 276・8. 6037-6045 (2001)
• [Publications] Mazaki, Y., Hashimoto, S., Okawa, K., Tsubouchi, A., Nakamura, K., Yagi, R., Yano, H., Kondo, A., Iwamatsu, A., Mizoguchi, A., Sabe, H.: "An ADP-Ribosylation Factor GTPase-activating Protein Git2-short/KIAA0148 Is Involved in Subcellular Localization of Paxillin and Actin Cytoskeletal Organization"Mol.Biol.Cell. 12・3. 645-662 (2001)
• [Publications] Uchida, H., Kondo, A., Yoshimura, Y., Mazaki, Y., Sabe, H.: "PAG3/PAPα/KIAA0400, a GTPase-activating protein for ADP-ribosylation factor, regulates ARF6 in FcγR-mediated phagocytosis of macrophages"J.Exp.Med.. 193・8. 955-966 (2001)
• [Publications] Nakamura, K., Yano, H., Schaefer, E., Sabe, H.: "Different modes and qualities of tyrosine phosphorylation of Fak and Pyk2 during epithelial-mesenchymal transdifferentiation and cell migration : Analysis of specific phosphorylation events using site-directed antibodies"Oncogene. 20・21. 2626-2635 (2001)
• [Publications] Yagi, R., Ishimaru, S., Yano, H., Gaul, U., Hanafusa, H., Sabe, H.: "A novel muscle LIM-only protein is generated from paxillin gene locus in Drosophila"EMBO Rep.. 2・9. 814-820 (2001)
• [Publications] Woods, AJ., Roberts, MS., Choudhary, J., Barry, ST., Mazaki, Y., Sabe, H., Morley, SJ., Critchley, DR., Norman, JC.: "Paxillin associates with poly(A)-binding protein 1 at the dense endoplasmic reticulum and the leading edge of migrating cells"J.Biol.Chem.. 277・8. 6428-6437 (2002)
• [Publications] 矢野 元, 佐邊 壽孝: "わかる実験医学シリーズ シグナル伝達がわかる・羊土社"細胞接着・細胞運動のしくみとシグナル伝達. 8 (2001)
• [Publications] A.Kondo,S.Hashimoto,H.Yano,K,Nagayama,Y.Mazaki & H.Sabe.: "A new paxillin-binding protein, PAG3/Papα/KIAA0400, bearing an Arf GTPaseactivating protein activity is involved in paxillin recruitment to focal adhesions and cell migration."Mol.Biol.Cell. 11・4. 1315-1327 (2000)
• [Publications] A.Ito,T.R.Kataoka,M.Watanabe,K.Nishiyama,Y.Mazaki,H.Sabe,Y.Kitamura & H.Nojima.: "A truncated isoform of PP2A promotes cell motility through paxillin phosphorylation."EMBO J.. 19・4. 562-571 (2000)
• [Publications] H.Yano,H.Uchida,T.Iwasaki,M.Mukai,H.Akedo,K.Nakamura,S.Hashimoto & H.Sabe.: "Paxillin α and Crk-associated substrate exert opposing effects on cell migration and contact inhibition of growth through tyrosine phosphorylation."Proc.Natl.Acad.Sci.USA. 97・16. 9076-9081 (2000)
• [Publications] T.Kanazawa,S.Nakamura,M.Momoi,T.Yamaji,H.Takematsu,H.Yano,H.Sabe,A.Yamamoto,T.Kawasaki & Y.Kozutsumi.: "Inhibition of cytokinesis by a lipid metabolite, psychosine."J.Cell Biol.. 149・4. 943-950 (2000)
• [Publications] K.Nakamura,H.Yano,H.Uchida,S.Hashimoto,E.Schaefer & H.Sabe.: "Tyrosine phosphorylation of paxillin is involved in temprospatial regulation of paxillin-containing focal adhesion formation and F-actin organization in motile cells."J.Biol.Chem.. 275・35. 27155-27164 (2000)
• [Publications] A.Suzuki,T.Hara,N.Kadota,Y.Nakagami,T.Izumi,T.Takenawa,H.Sabe & T.Endo.: "Meltrin a cytoplasmic domain interacts with SH3 domains of Src and Grb2 and is phosphorylated by Src."Oncogene. 19・51. 5842-5850 (2000)
• [Publications] Y.Ishii,A.Ochiai,T.Yamada,S.Akimoto,K.Yanagihara,M.Kitajima,& S.Hirohashi.: "Integrin α6β4 as a suppressor and a predictive marker for peritoneal dissemination in human gastric cancer."Gastroenterology. 118・3. 497-506 (2000)
• [Publications] T.Genda,M.Sakamoto,T.Ichida,H.Asakura,& S.Hirohashi.: "Loss of cell-cell contact is induced by integrin-mediated cell-substratum adhesion in highly-motile and highly-metastatic hepatocellular carcinoma cells."Lab Invest. 80・3. 387-394 (2001)
• [Publications] S.Hashimoto,A.Tsubouchi,Y.Mazaki & H.Sabe.: "Interaction of paxillin with p21-activated kinase (PAK) : association of paxillin α with the kinase-inactive and the Cdc42-activated forms of PAK3."J.Biol.Chem.. (in press)..
• [Publications] Y.Mazaki,S.Hashimoto,K.Okawa.K.Nakamura,R.Yagi,H.Yano,A.Kondo,A.Iwamatsu,A.Mizoguchi,& H.Sabe.: "An ARFGAP protein Git2-short/KIAA0148 Is Involved in Subcellular Localization of Paxillin, and Actin Cytoskeletal Organization."Mol.Biol.Cell. (in press)..
• [Publications] K.Nakamura,H.Yano,E.Schaefer & H.Sabe.: "Different modes and qualties of tyrosine phosphorylation of Fak and Pyk2 during epithelial-mesenchymal transdifferentiation and cell migration."Oncogene. (in press)..
• [Publications] M.takamura,M.Sakamoto,T.Genda,T.Ichida,H.Asakura,& S.Hirohashi.: "Inhibition of intrahepatic metastasis of human hepatocellular carcinoma by Rhoassociated protein kinase inhibitor Y-27632."Hepatology.. (in press).
• [Publications] 佐邊壽孝: "岩波講座、現代医学の基礎2"分子・細胞の生物学II""細胞-細胞、細胞-基質の相互作用""編集:石川春律、高井義美、月田承一郎 出版社:岩波書店. 27 (2000)