遺伝子組換えHGFレセプターアンタゴニストによる癌細胞の浸潤・転移抑制剤の開発

1995 Fiscal Year Annual Research Report

• Project/Area Number: 07557199
• Research Category: Grant-in-Aid for Developmental Scientific Research (B)
• Research Institution: Osaka University
• Principal Investigator: 中村 敏一 大阪大学, 医学部, 教授 (00049397)
• Co-Investigator(Kenkyū-buntansha): 河田 純男 大阪大学, 医学部, 助教授 (90183285) ; 門田 守人 大阪大学, 医学部, 教授 (00127309) ; 根本 康夫 大阪大学, 医学部, 助手 (30250088) ; 松本 邦夫 大阪大学, 医学部, 講師 (90201780)
• Keywords: HGF / c-met / 癌細胞 / 腫瘍 / 癌浸潤・転移 / 増殖因子 / サイトカイン / レセプター

Research Abstract

本年度の主な研究成果は以下の通りである.
1)上皮系癌細胞(carcinoma)の多くはHGFレセプターであるc-met遺伝子産物を発現していることが知られている.これら癌細胞に対するHGFの作用を調べたところ,HGFはin vitroにおいて肺癌細胞や胆管上皮癌細胞,胆嚢癌細胞などの増殖を促進させるのみならず,コラーゲンゲルやマトリゲル層への浸潤・遊走を促すことが明らかになった.またこれらの癌細胞においてはHGFによる培養血管内皮細胞への接着能促進効果も認められた.
2)多くの上皮系癌細胞は間質線維芽細胞に対してHGF産生を促進させる誘導因子(インジュリン)を産生している.これらの因子を同定したところ,癌細胞由来インジュリン活性は主にlL-1,bFGF,PDGFなどであることが明らかになった.しかしある種のマウス乳癌由来細胞株はこれら既殖因子・サイトカインとは異なる低分子のHGF誘導因子を産生していることを見い出し,現在,その分子の単離・構造決定を進めている.
3)HGFによりその増殖ならびに浸潤・転移能が促進する癌細胞の悪性化を抑制することを目的にHGFレセプター・アンタゴニストの探索・開発を試み,HGFの限定分解産物がアンタゴニスト活性を有することを明らかにした.HGFa鎖の4つのクリングル・ドメインを有するこのアンタゴニスト分子はHGFのc-MET/HGFレセプターへの結合を阻害し,in vitroにおけるHGFの増殖促進活性のみならずコラーゲンゲルやマトリゲル層内への浸潤をほぼ完全に阻害することが確認された.

Research Products

• [Publications] N.Nakayama: "Differing distribution of hepatocyte growth factor-positive cells in the liver of LEC rats with acute hepatitis,chronic hepatitis and hepatoma." Jap.J.Cancer Res.86. 5-9 (1995)
• [Publications] H.Nakamura: "Molecular cloning of Xenopus HGF cDNA,and its expression studied in Xenopus early embryogenesis." Mech.Develop.49. 121-131 (1995)
• [Publications] G.Shiota: "Serum hepatocyte growth factor level:its clinical implication." Hepatology. 21. 106-112 (1995)
• [Publications] W.G.Jiang: "Inhibition of motility and invasion by invasion inhibiting factor-2(11F-2)on human colon cancer cells." Surg.Res.Commun.17. 67-78 (1995)
• [Publications] K.Matsumoto: "Novel function of prostaglandins as inducer of gene expression of hepatocyte growth factor." J.Biochem.117. 458-464 (1995)
• [Publications] F.Myokai: "Expression of the hepatocyte growth factor gene during development of the chicken limb." Develop.dynamics. 202. 80-90 (1995)
• [Publications] T.Sato: "Hepatocyte growth factor(HGF)is involved in the formation of osteoclast-like cells mediated by clonal stromal cells(MC3T3-G1/PA6)." J.Cell.Physiol.164. 197-204 (1995)
• [Publications] M.J.Gomez-Lechon: "Effect of hepatocyte growth factor(h-rHGF)on the growth and metabolism of human hepatocytes in primary culture." Hepatology. 21. 1248-1254 (1995)
• [Publications] S.Saito: "Hepatocyte growth factor promotes the growth of cytotrophoblasts by the paracrine system." J.Biochem.117. 671-676 (1995)
• [Publications] M.Takahashi: "Hepatocyte growth factor is the most potent endogenous stimulant of rabbit gastric epithelial cell proliferation and migration in primary culture." J.Clin.Invest.95. 1994-2003 (1995)
• [Publications] T.Takabayashi: "Hepatocyte growth factor/scatter factor has mitogenic,motogenic and morphogenetic activities on chondrocytes." J.Cell Biol.129. 1411-1419 (1995)
• [Publications] K.Matsumoto: "Hepatocyte growth factor as a potent survival factor for rat pheochromocytoma PC12 cells." Exp.Cell Res.220. 71-78 (1995)
• [Publications] S.Seslar: "Tumor-stroma interactions and stromal cell density regulate hepatocyte growth factor protein levels:A role for transforming growth factor-b activation." Endocrinology. 136. 1945-1953 (1995)
• [Publications] A.Yamada: "Rapid and sensitive enzyme-linked immunosorbent assay for measurement of HGF in rat and human tissues." Biomed.Res.16. 105-114 (1995)
• [Publications] E.J.Barros: "Differential tubulogenic and branching morphogenetic activities of growth factors:Implications for epithelial tissue development." Proc.Natl.Acad.Sci.USA. 92. 4412-4416 (1995)
• [Publications] H.Shimura: "Induction of invasive growth in a gallbladder cancer cell line by hepatocyte growth factor in vitro." Jap.J.Cancer Res.86. 662-669 (1995)
• [Publications] K.Ohashi: "Molecular cloning,characterization and in situe localization in the brain of rat sky receptor tyrosine kinase." J.Biochem.117. 1267-1275 (1995)
• [Publications] T.Adachi: "Roles of prostaglandin production and MAP kinase activation in hepatocyte growth factor-mediated rat hepatocyte proliferation." Hepatology. 21. 1668-1674 (1995)
• [Publications] M.Yaono: "Hepatocyte growth factor enhancement of GST-P positive hepatic foci development in rats treated with diethylnitrosamine and N-ethyl-N-hydroxy ethylnitrosamine." Jap.J.Cancer Res.86. 718-723 (1995)
• [Publications] W.G.Jiang: "Inhibition of HGF/SF-induced membrane ruffling and cell motility by transient elevation of cytosolic free Ca^<2+>." Exp.Cell Res.220. 424-433 (1995)
• [Publications] Y.Matsuda: "Hepatocyte growth factor prevents liver cirrhosis caused by dimethylnitrosamine in rats." J.Biochem.118. 643-649 (1995)
• [Publications] S.Honda: "Localization and functional coupling of HGF and c-Met/HGF receptor inrat brain:implication as neurotrophic factor." Mole.Brain Res.32. 197-210 (1995)
• [Publications] M.R.Vila: "Hepatocyte growth factor is a potent mitogen for normal human pancreas cells in vitro." Lab.Invest.73. 409-418 (1995)
• [Publications] P.Gines: "Tyrosine kinase growth factor receptors but not seven-membrane-spanning or phorbol esters activate mitogen-activated protein kinase in rat hepatocytes." Hepatology. 22. 1296-1303 (1995)
• [Publications] M.Hu: "Human peri-tumoral and lung fibroblasts produce paracrine motility factors for recently established human sarcoma cell strains." Intn.J.Cancer. 62. 585-592 (1995)
• [Publications] Y.Nakamura: "Expression of local hepatocyte growth factor system in vascular tissue." Biochem.Biophys.Res.Commun.215. 483-488 (1995)
• [Publications] M.Takahashi: "Hepatocyte growth factor stimulates wound repair of the rabbit esophageal epithelial cells in primary culture." Biochem.Biophys.Res.Commun.216. 298-305 (1995)
• [Publications] W.G.Jiang: "Induction of tyrosine phosphorylation and translocation of ezrin by hepatocyte growth factor/scatter factor(HGF/SF)." Biochem.Biophys.Res.Commun.217. 1062-1069 (1995)
• [Publications] T.Kaido: "Continous HGF supply from spleen transplanted HGF-expressing fibroblasts prevents CCl_4-induced acute liver injury in rats." Biochem.Biophys.Res.Commun.218. 1-5 (1996)
• [Publications] K.Matsumoto: "Hepatocyte growth factor as a renotropic factor and therapeutic potential for acute renal failure." Acute Renal Failure:Emerging New Concepts and Therapeutic Strategies., 24 (1995)
• [Publications] H.Amaike: "Takahashi Memorial Forum,Tokyo,5 November 1994,Excepta Medica International Congress Series 1086" Elsevier Science B.V.Amsterdam., 14 (1995)
• [Publications] 坂倉照好: "細胞外マトリックス" 羊土社, 13 (1995)
• [Publications] 中村敏一: "用語ライブラリー:発生・神経" 羊土社, 163 (1996)
• [Publications] 中村敏一 監訳: "細胞増殖因子(I.A.McKay and I.M.Leigh編)" メディカル・サイエンス インターナショナル, 232 (1995)
• [Publications] U.Koshimizu: "Liver and Environmental Xenobiotics" Narosa-Elsevier Verlag(in press), (1996)