上皮・間葉相互作用によるがん悪性化の分子機構とその制御

2000 年度 実績報告書

• 研究課題/領域番号: 12215082
• 研究種目: 特定領域研究(C)
• 研究機関: 大阪大学
• 研究代表者: 中村 敏一 大阪大学, 医学系研究科, 教授 (00049397)
• 研究分担者: 小清水 右一 大阪大学, 医学系研究科, 助手 (50281126) ; 船越 洋 大阪大学, 医学系研究科, 助手 (40273685) ; 松本 邦夫 大阪大学, 医学系研究科, 助教授 (90201780)
• キーワード: HGF / NK4 / HGFアンタゴニスト / 血管新生 / 浸潤・転移 / クリングルドメイン / LIMキナーゼ / 細胞骨格

研究概要

1)NK4による血管新生抑制機構の解明
NK4はN末端ヘアピンドメインと4個のクリングルドメインをもつHGFの分子内断片である。NK4の分子内N末端ヘアピンドメインがc-Metレセプターに結合する上で必須のドメインであり、このドメインを欠く4個のクリングルドメイン(K1-4)はHGFアンタゴニスト活性を失うことが予想された。そこでリコンビナントNK4ならびにK1-4を調製し、両分子のHGFアンタゴニスト活性ならびに血管新生阻害活性を調べた。その結果、NK4はHGFによる肝細胞の増殖促進活性を阻害し、HGFアンタゴニスト活性を示したが、K1-4はHGFアンタゴニスト活性を失っていた。一方、培養ヒト血管内皮細胞に対するNK4ならびにK1-4の作用を調べた結果、K1-4はNK4と同等にbFGF、VEGF、HGFによって促進されるヒト血管内皮細胞の増殖ならびに遊走を阻害した。したがってNK4のHGFアンタゴニスト活性は血管新生阻害作用に関与せず、血管新生阻害作用には分子内クリングルドメインが関与することが明らかになった。
2)LIMキナーゼの活性調節機構および生体機能の解析
LIMキナーゼ(LIMK)はコフィリンを特異的にリン酸化するアクチン細胞骨格の制御因子である。私達はこれまでにLIMK1、LIMK2が異なるRhoファミリー分子の制御下で活性化されること、すなわちLIMK1はRacにより、一方、LIMK2はRhoおよびCdc42により活性化されることを報告しているが、さらに今回、LIMK2はRho依存性キナーゼであるROCKによるリン酸化を受け、活性化されることを明らかにした。またXenopus卵において、LIMKは細胞(減数)分裂の際に重要な役割を担っている微小管形成中心の形成に関与していること、さらにLIMK2ノックアウトマウスではその精子形成に異常が認められることを見い出した。

研究成果

• [文献書誌] S.Hiscox: "Inhibition of HGF/SF-induced breast cancer cell matility and invasion by the HGF/SF variant, NK4."Breast Cancer Res.& Treat.. 59. 245-254 (2000)
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• [文献書誌] T.Takahashi: "Functional involvement of LIM-kinase in Xenopus oocyte maturation."Develop.Biol.. 229. 554-567 (2001)
• [文献書誌] T.Sumi: "Specific activation of LIM-kinase 2 via threonine-505 phosphorylation by ROCK under the control of Rho."J.Biol.Chem.. 276. 670-676 (2001)
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