HGFならびにLIMキナーゼによる高次細胞機能の調節とがん特性の研究

1997 Fiscal Year Annual Research Report

• Project/Area Number: 06283216
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Research Institution: Osaka University
• Principal Investigator: 中村 敏一 大阪大学, 医学部, 教授 (00049397)
• Co-Investigator(Kenkyū-buntansha): 宮本 一政 大阪大学, 医学部, 助手 (60273679) ; 小清水 右一 大阪大学, 医学部, 助手 (50281126) ; 松本 邦夫 大阪大学, 医学部, 助教授 (90201780)
• Keywords: HGF / HGFアンタゴニスト / 腫瘍血管新生 / 脂肪肝 / 慢性腎不全 / 劇症肝炎 / LIMキナーゼ / Sky

Research Abstract

今年度の主な研究成果を以下に記す。
1.HGFアンタゴニストによる浸潤・転移、腫瘍血管新生の阻止:HGFアンタゴニスト(HGF/NK4)の調製に成功した。HGFは様々ながん細胞に対しin vitroにおける浸潤を強く促進したが、HGF/NK4はこのin vitro浸潤をほぼ完全に阻害した。さらにがん細胞をマウス皮下に移植しHGF/NK4をがん組織近傍に注入した結果in vivoでの増殖、浸潤、腫瘍血管新生を抑制し、さらにLewis肺がんならびにJyg乳がんの肺転移をも抑制した。HGF/NK4はがんの浸潤・転移・血管新生を抑える新しい制癌剤となると考えられる。
2.HGFによる劇症肝炎の防止ならびに脂肪肝の改善・治癒作用:HGFは抗Fas抗体による大量肝細胞アポトーシスならびに劇症肝炎の発症をほぼ完全に抑制し、同様の結果はエンドトキシンで誘起した劇症肝炎モデルでも認められた。両モデルにおいてHGFはBcl-xlの発現を著明に誘導し、この作用がHGFによる強力な抗アポトーシス作用の1つの機作と考えられた。また、HGFはエタノール摂取によって引き起こされた脂肪肝に対し著しい改善・治癒作用を示した。HGFは劇症肝炎に対する発症阻止ならびに肝再生促進能をもつ有効な治療薬となるものと考えられる。
3.HGFからみた慢性腎不全の発症機構とHGFによる治癒・改善作用:ヒトの慢性腎不全と同様の病態を呈するICGNマウスは生後2〜3ヵ月でネフローゼや腎線維化が認められるが、それに伴いTGF-βの産生レベルは増加し、HGFレベルは著しく減少した。内因性HGFレベルの低下を補うべくHGFを4週間投与したところ、TGF-βのレベルは低下し、腎線維化は著しく軽減された。さらにHGFは尿細管上皮細胞のアポトーシスを強く抑制する一方、尿細管の再構築を促進し、腎機能はほぽ正常レベルまで改善された。HGFは根本的治療法のない慢性腎不全に対して有効な治療薬となるものと思われる。
4.LIMキナーゼ(LIMK)相互作用分子の同定:LIMK1ならびにLIMK2の相互作用分子をtwo hybrid法で約56種類同定した。そのうち新規分子は24種類であり。細胞骨格、細胞周期、転写調節に関係する分子と考えられた。また精巣において特異的に発現するLIMK2バリアント(LIMドメイン欠損型)を発見した。
5.脳神経特異的レセプター型キナーゼSkyの機能:Skyと、そのリガンドGas6の脳神経系に於ける役割を解析した結果、Gas6がSkyを介して大脳皮質、海馬などの多くの神経細胞の生存を促す新しいneurotrophic factorとして機能することを見い出した。

Research Products

• [Publications] K.Takai, et al.: "Hipatocyte growth factor is constitutively produced by human bone marrow stromal cells and promotes erythropoiesis." Blood. 89. 1560-1565 (1997)
• [Publications] B.Morishita, et al.: "Potential role of endothelium-specific growth factor hepatocyte growth factor,on endothelial damage in diabetes militus." Diabetes. 46. 138-142 (1997)
• [Publications] I.S.Weimar, et al.: "Hepatocyte growth factor/scatter factor(HGF/SF)promotes adhesion of lymphoma cells to extra-cellular matrix molecules via a4b,and a5b,integrens." Blood. 89. 990-1000 (1997)
• [Publications] N.Kawakami, et al.: "Involvement of hepatocyte growth factor in increased integrin expression on Hep G2 cells triggered by adhesion to endothelial cells." Br.J.Cancer. 75. 47-53 (1997)
• [Publications] T.Sakai, et al.: "Hepatocyte growth factor in bronchoalveolar lavage fluid and cells in patients with in flammatory chest diseases of the lower respitatory tract:Defection by RIA and in situ hybridization." Am.J.Respir.Cell Mole Biol.16. 388-397 (1997)
• [Publications] T.Takahashi, et al.: "Xenopus LIM motif-containing protein kinase,XLIMK,is expressed in the developing head structure of the embryo." Develop.Dyn.209. 196-205 (1997)
• [Publications] S.Aoki, et al.: "Activation of Met tyrosine kinase by hepatocyte growth factor is essential for internal ganogenesis in Xenopus embryo." Biochem.Biophys.Res.Commun.234. 8-14 (1997)
• [Publications] Y.Ikeda, et al.: "Bile ductular proliferation in the granulation tissue of rat liver.:Histological and serial reconstruction studies." Biomed.Res.18. 113-127 (1997)
• [Publications] Y.Matsuda, et al.: "Prevential and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis." Hepatology. 26. 81-89 (1997)
• [Publications] K.Furugori, et al.: "Level of Hepatocyte growth factor and its messenger ribonucleic acid in uncomplecated pregnancies and those complicated by preeclamptia." J.Clin.Endocrinol.& Metabo.82. 2726-2730 (1997)
• [Publications] T.Nakamura, et al.: "Induction of hepatocyte growth factor in fibroblasts by tumor-derived factors affects invasive growth of tumor cells:In vitro analysis of tumor-stromal interaction." Cancer Res.57. 3305-3313 (1997)
• [Publications] T.Imai, et al.: "Sparse distribution of hepatocyte growth factor-producing cells inside hepatocellular foci in rats treated with hepatocarcinogens." Jpn.J.Cancer Res.88. 615-618 (1997)
• [Publications] T.Kaido, et al.: "Continous hepatocyte growth factor supply prevents lipopolysacharide-induced liver injury in rats." FEBS Lett.411. 378-382 (1997)
• [Publications] S.E.Hiscox, et al.: "Expresstion of the HGF/SF receptor c-met,and its ligand in human colon cancers." Cancer Investigations. 15. 513-521 (1997)
• [Publications] R.Morishita, et al.: "Role of hepatocyte growth factor in endothelial regulation:Prevention of high D-glucose induced endothelia lcell death by prostglandins and phosphodiesterase type 3 inhibitor." Diabetologia. 40. 1053-1061 (1997)
• [Publications] H.Li, et al.: "Hepatocyte growth factor stemuolates invasion of gallbladder carcinoma cell lines in vitro." Clin.Exp.Metastasis.15. 484-493 (1997)
• [Publications] K.Liu, et al.: "Existence of two nonliner elimination mechanisms for hepatocyte growth factor in rats." Am.J.Physiol. 273. E891-E897 (1997)
• [Publications] S.Wakitani, et al.: "Hepatocyte growth factor faulitates Cartilage repair:full thickness articular cartilage defect studied in rabbit knee." Acta Orthop.Stand.68. 474-480 (1997)
• [Publications] T.Adachi, et al.: "Possible involvement of pertussis toxin-sensitive G protein in hepatocyte growth factor(HGF)-induced signal transduction in cultured rat hepatocytes." Hepatology.26. 295-300 (1997)
• [Publications] V.R.Wallenius, et al.: "Chromosomal localization of rat hepatocyte growth factor (Hgf)and HGF receptor (Met) and chractrization of HGF receptor cDNA." Mammalian Genome. 8. 661-667 (1997)
• [Publications] N.Nakano, et al.: "Role of antagotensin II in the reguration of a novel vascular modulator,hepatocyte growth factor(HGF),in experimental hypertensive rats." Hypertension. 30. 1448-1454 (1997)
• [Publications] K.Date, et al.: "HGF/NK4 is a specific antagonist for pleiotrophic actions of hepatocyte growth factor." FEBS Lett.420. 1-6 (1997)
• [Publications] A.Schmassman, et al.: "Role of antagotensin II in the reguration of a novel vascular modulator,hepatocyte growth factor(HGF),in experimental hypertensive rats." Gastroenterolgy. 113. 1858-1872 (1997)
• [Publications] U.Koshimizu, et al.: "cDNA cloning,genomic organization and chromosomal localization of the mouse Lim motif-containing kinase,Limk 2 gene." Biochem.Biophys.Res.Commun.241. 243-250 (1997)
• [Publications] M.Yaekashiwa, et al.: "Both simultaneous and delayed administration of hepatocyte growth factor(HGF)repress the fibrotic changes in murine lung injury induced by bleomycin." Am.J.Resp. & Clin.Care Med.156. 1937-1944 (1997)
• [Publications] R.Morishita, et al.: "Potential role of a novel vascular modulator,hepatocyte growth factor(HGF),in cardiovascular disease:characterization and regulation of local HGF system." J.Atherosclerosis and Thrombosis. 4. 12-19 (1997)
• [Publications] U.Koshimizu, et al.: "Hepatotrophic activities of HGF in liver regeneration after injury and the clnical potentiality for liver diseases." “Liver and Environmental Xenobiotics" Narosa-Elsevier Verlag,. 220-229 (1997)
• [Publications] K.Matsumoto, et al.: "HGF :its organotrophic role and therapeutic potential." “Plasminogen-Related Growth Factors". 212. 198-214 (1997)
• [Publications] K.Matsumoto, et al.: "Hepatocyte growth factor(HGF)as a tissue organizer for organogenesis and regeneration." Biochem.Biophys.Res.Commun.(Mini-Review). 239. 639-644 (1997)
• [Publications] K.Matsumoto, et al.: "Pathophysiological aspects and therapeutic potential of hepatocyte growth factor in liver disease." Internal Medicine and Endocrinology:Pathophysiological and clinical interactions(eds by A.Fabbrini and V.Santiemma.). 95-106 (1997)
• [Publications] D.Schuppan, et al.: "Collagens in the liver extracellular matrix hepatocyte growth factor." Gastroenterology.114. 139-152 (1998)
• [Publications] M.Lyon, et al.: "Hepatocyte growth factor/scatter factor binds with high affinity to dermatan sulphate." J.Biol.Chem.273. 271-278 (1998)
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• [Publications] K.Liu et al.: "Protamine enhances the proliferative activity of hepatocyte growth factor in rats." Am.J.Physiol.(in press). (1997)
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• [Publications] K.Liu et al.: "Change in hepatic handling of hepatocyte growth factor duing liver regeneration in rats." Am.J.Physiol.269(in press). (1998)
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• [Publications] H.Ohmichi et al.: "Hepatocyte growth factor(HGF)acts as a mesenchymal-derived morphogenic factor during fetal lung devel pment." Development.(in press). (1998)
• [Publications] T.Miyazawa, et al.: "Protection of hippocampal neurons from ischemia-induced delayed neuronal death by hepatocyte growth factor:A novel neurotrophic factor." J.Celebral Blood Flow Metabolism.(in press). (1998)
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• [Publications] F.Tamura, et al.: "FK506 promotes liver regeneration by suppressing natural-killer cell activity.natural killer cell activity." J.Gastroenterol.and Hepatol.(in press). (1998)
• [Publications] W.G.Jiang, et al.: "Hepatocyte growth factor/scatter factor,its molecular,cellular and clinical impacts in cancer." Crit.Review Onclo.Hematol.(in press). (1998)
• [Publications] 中村 敏一: "標準分子医化学" 医学書院, 843-852 (1997)
• [Publications] 中村 敏一: "分子生物学プロトコール改訂版 タンパク質の生化学的分析" 南江堂 (印刷中), (1997)
• [Publications] 中村 敏一: "肝臓病-今日の診断と最新の治療" 国際医書出版, 299-310 (1997)
• [Publications] 中村 敏一: "生命体システムの修復" 講談社サイエンティフィック, 124 (1997)
• [Publications] 松本 邦夫, 中村 敏一: "病理学キーワード" 文光社, pp.40 (1997)