1992 Fiscal Year Annual Research Report

HGF(肝細胞増殖因子)の大量発現法ならびに高感度イムノアッセイ法の確立

Research Project

Project/Area Number	03558020
Research Institution	Osaka Univrsity
Principal Investigator	中村敏一大阪大学, 医学部, 教授 (00049397)
Co-Investigator(Kenkyū-buntansha)	竹原豊浩東洋紡績(株), 医薬研, 主任部員萩屋道雄東洋紡績(株), 医薬研, 主席部員松本邦夫九州大学, 理学部, 助手 (90201780) 原野友之九州大学, 理学部, 助手 (80037275) 水野健作九州大学, 理学部, 助教授 (70128396)
Keywords	HGF / 肝再生 / 腎再生 / 抗肝炎作用 / ELISA / RIA
Research Abstract	HGF(肝細胞増殖因子)は成熟肝細胞をはじめ多くの上皮細胞を標的とする増殖因子で、1989年私達はHGFのcDNAクローニングに成功し、世界に先駆けて全一次構造を解明した。HGFは発見当初から実体の不明であった肝再生因子の本体であると考えられてきたが、私達はHGFが賢再生因子としても機能することを見い出した。本研究はHGFの大量発現法ならびに高感度イムノアッセイ法の確立を主たる目的として行なわれたが平成4年度においては以下の成果が得られた。まず、遺伝子工学的手法を用いてラットHGFの大量発現法を開発し、ラットHGFに対するポリクローナル抗体を作製し、ラットHGFの高感度ELISA系を確立した。また、実験的に様々な肝傷害あるいは賢傷害を引き起こしたマウスにヒトリコンビナントHGFを尾静脈より投与したところ、HGFが肝再生ならびに賢再生を著しく促進することが明らかになり、HGFがin vivoで肝再生因子、賢再生因子として機能することを実証した。しかもHGFはα-naphtylisothio-cyanateによって引き起こされる急性の肝炎を抑え、肝機能を維持する抗肝炎作用があることが明らかになった。また、HGFによる肝再生の促進作用をビーグル犬を用いた実験系においても実証した。次に本研究において確立したHGFの高感度ELISA法を用いて、肝炎、肝癌切除、賢癌、急性腎不全、肺炎、肺癌などの患者血清中のHGFレベルを測定したところ、これら種々の臓器疾患においてHGFレベルが著しく増加していることが明らかになった。以上の研究成果はいずれもHGFが種々の肝疾患、腎疾患さらには胚疾患等に対する有効な治療薬、診断薬となりうることを示している。今後、各種臓器疾患モデル動物を用いたHGFの投与実験、病態予後の判定を目的としたHGFレベルの詳細な検討を行なうことによって、HGFが治療薬、診断薬として利用され、臨床医学の分野に多大な貢献をするものと考えられる。

Research Products
(18 results)

All Other

All Publications (18 results)

[Publications] G.Shiora,D.B.Rhoads,T.C.Wang, T.Nakamura and E.V. Schmidt: "Hepatocyre growth factor inhibits growth of hipatocellular carcinoma cells." Proc.Matl.Acad.Sci.USA. 89. 373-377 (1992)
[Publications] H. Tajuma,O. Higuchi,K. Mizuno and T.Nakamura: "Tissue distribution of hepatocyre growth factor receptor and its exclusive down-regulation in a regenerating organ afterinjury." J.Biochem.111. 401-406 (1992)
[Publications] K.Yanagita,M.Nagaike,H. Ishibashi, Y. Niho, K. matsumoto and T.Nakamura: "Lung may have an endocrine function producing hepatocyte growth factor in re-sponse to injury of distal orgnas." Biochem.Biophys.Res. Commun.182. 802-809 (1992)
[Publications] K.Matsumoto,H.Tajuma,M.Hamanoue, S. Kono, T.Kinoshita and T.nakamura: "Identification and characterization of ¨injurin¨.an inducer of expression of the gene for hepatocyte growth factor." Proc.Natl.Acad.Sci.USA. 89. 3800-3804 (1992)
[Publications] O.Higuchi,K.Mizuno,G.F.Vande Woude and T.Nakamur: "Expression of the c-met proto-oncogene in COS cells es required for hepatocyte growth factor high affinity binding and mitogenic signal trnsduction." FEBS Lett.301. 282-286 (1992)
[Publications] M.Hamanoue,K.Kawaida,S.Takao,H.Shimazu,S.Noji,K.Matsumoto and T.Nakamura: "Rapid and marked induction of HGF mRNA in liver and HGF activity in serum of rats after ischemic of crush liver injury." Hepatology. 16. 1485-1492 (1992)
[Publications] H.Tajima.K.Matsumoto and T.Nakamura: "Regulation of cell growth and motility by hepatocyte growth factor and receptorex-prssion in various cell species." Exp.Cell Ees.202. 423-431 (1992)
[Publications] S. Kono, M. Nagaike, K. Matsumoto,and T.Nakamura: "Marked induction of hepatocyte growth factor mRNA in intact kidney and spleen in response to injury of distan organ." Biochem.Biophys.Res.Commun.186. 991-998 (1992)
[Publications] Y.shiki,H.Ohmishi,Y.Muto,K.Matsumoto and T.Nakamura: "Direct evudence that heparocyt growth factor es a hepatotrophic factor for liver regeneration and for potent anti-hepatitis action in vivo." Hepatology. 16. 1227-1235 (1992)
[Publications] K.Matsumoto,H.Tajima,H.Okazaki,and T.Nakamura: "Negative regulation of hepatocytr growth factor gene expression in human lung fibroblasts and leukemic cells by trnsforming growth factor-β I and glucocorti-coids" J.Biol.Chem.267. 24917-24920 (1992)
[Publications] K.Matsumoto,H.Okazaki,and T.Nakamura: "Up-regulation of hepatocyte growth factor gene exprssion by interleukin-l in human skun febroblasts." Biochem.Biohpys.Commun.189. 235-243 (1992)
[Publications] K.Matsumoto and T.Nakamura: "Hepatocyte growth factor:Molecular stucture,roles in liver regeneneration,and other biological functions." Criteal Reviews in Oncogenesis. 3. 27-54 (1992)
[Publications] K.Mizuno.T.Takehara and T.Nakamura: "Proteolytic activation of shigle-chain precursor of hepatocyte growth factor by ex-tracellular serine protease." Biochem.Biophys,Res.Commun.189. 1631-1638 (1993)
[Publications] G.Shiota,T.C.Wang,S.dela Monte,T.Nakamura,and E.V.Schmidt: "Hepatocyte gorwth factor accelerates liver regeneration and causes a hepato-prolif-erative disorder in transgenic mice." J.Clin.Invest.(1993)
[Publications] K.Mizuno and T.Nakamura(ed.I.D.Goddbergの分担): "Molecilar characteristics of HGF and gene,and its biochemical aspects.In “Hepa-tocyte Growth Factor Factor-Scatter Factor and c-Met Eeceptor"" Birkhauser Verlag, 1-29 (1993)
[Publications] K.Matsumoto and T.Nakamura(ed.I.D.Goldbergの分担): "Roles of HGF as a pleiotrpic factor in organ eegeneeration.In“Hepatocyte Growth Factor-Scatter Aactor and c-Met Receptot"" Birkhauser Verlag, PP.225-249 (1993)
[Publications] K.Matsumoto T.Nakamura(eds.Drs.J.B.Griffiths,A.Doyle and D.G.Newellの分担): "Primary and continuous cell culture:Collagenase and Hyaluronidase.In “Protocols in Cell and Tissue Culture"" john Wiley & Sone Limited,England, (1993)
[Publications] T.Nakamura,K.Muzuno,and K.Matsumoto(eds.T.Tsuruta,M.Seno and M.Doyamaの分担): "Hepatocyte growth factor:Mitogen,motogen,morphpgen,and its roles in tissue regeneration In “New Functionalty Materials:Design,Preparation and Control." Elsevier Science Prblishers B.V., (1993)

1992 Fiscal Year Annual Research Report

HGF(肝細胞増殖因子)の大量発現法ならびに高感度イムノアッセイ法の確立

Principal Investigator

中村 敏一 大阪大学, 医学部, 教授 (00049397)

Research Products

[Publications] G.Shiora,D.B.Rhoads,T.C.Wang, T.Nakamura and E.V. Schmidt: "Hepatocyre growth factor inhibits growth of hipatocellular carcinoma cells." Proc.Matl.Acad.Sci.USA. 89. 373-377 (1992)

[Publications] H. Tajuma,O. Higuchi,K. Mizuno and T.Nakamura: "Tissue distribution of hepatocyre growth factor receptor and its exclusive down-regulation in a regenerating organ afterinjury." J.Biochem.111. 401-406 (1992)

[Publications] K.Yanagita,M.Nagaike,H. Ishibashi, Y. Niho, K. matsumoto and T.Nakamura: "Lung may have an endocrine function producing hepatocyte growth factor in re-sponse to injury of distal orgnas." Biochem.Biophys.Res. Commun.182. 802-809 (1992)

[Publications] K.Matsumoto,H.Tajuma,M.Hamanoue, S. Kono, T.Kinoshita and T.nakamura: "Identification and characterization of ¨injurin¨.an inducer of expression of the gene for hepatocyte growth factor." Proc.Natl.Acad.Sci.USA. 89. 3800-3804 (1992)

[Publications] O.Higuchi,K.Mizuno,G.F.Vande Woude and T.Nakamur: "Expression of the c-met proto-oncogene in COS cells es required for hepatocyte growth factor high affinity binding and mitogenic signal trnsduction." FEBS Lett.301. 282-286 (1992)

[Publications] M.Hamanoue,K.Kawaida,S.Takao,H.Shimazu,S.Noji,K.Matsumoto and T.Nakamura: "Rapid and marked induction of HGF mRNA in liver and HGF activity in serum of rats after ischemic of crush liver injury." Hepatology. 16. 1485-1492 (1992)

[Publications] H.Tajima.K.Matsumoto and T.Nakamura: "Regulation of cell growth and motility by hepatocyte growth factor and receptorex-prssion in various cell species." Exp.Cell Ees.202. 423-431 (1992)

[Publications] S. Kono, M. Nagaike, K. Matsumoto,and T.Nakamura: "Marked induction of hepatocyte growth factor mRNA in intact kidney and spleen in response to injury of distan organ." Biochem.Biophys.Res.Commun.186. 991-998 (1992)

[Publications] Y.shiki,H.Ohmishi,Y.Muto,K.Matsumoto and T.Nakamura: "Direct evudence that heparocyt growth factor es a hepatotrophic factor for liver regeneration and for potent anti-hepatitis action in vivo." Hepatology. 16. 1227-1235 (1992)

[Publications] K.Matsumoto,H.Tajima,H.Okazaki,and T.Nakamura: "Negative regulation of hepatocytr growth factor gene expression in human lung fibroblasts and leukemic cells by trnsforming growth factor-β I and glucocorti-coids" J.Biol.Chem.267. 24917-24920 (1992)

[Publications] K.Matsumoto,H.Okazaki,and T.Nakamura: "Up-regulation of hepatocyte growth factor gene exprssion by interleukin-l in human skun febroblasts." Biochem.Biohpys.Commun.189. 235-243 (1992)

[Publications] K.Matsumoto and T.Nakamura: "Hepatocyte growth factor:Molecular stucture,roles in liver regeneneration,and other biological functions." Criteal Reviews in Oncogenesis. 3. 27-54 (1992)

[Publications] K.Mizuno.T.Takehara and T.Nakamura: "Proteolytic activation of shigle-chain precursor of hepatocyte growth factor by ex-tracellular serine protease." Biochem.Biophys,Res.Commun.189. 1631-1638 (1993)

[Publications] G.Shiota,T.C.Wang,S.dela Monte,T.Nakamura,and E.V.Schmidt: "Hepatocyte gorwth factor accelerates liver regeneration and causes a hepato-prolif-erative disorder in transgenic mice." J.Clin.Invest.(1993)

[Publications] K.Mizuno and T.Nakamura(ed.I.D.Goddbergの分担): "Molecilar characteristics of HGF and gene,and its biochemical aspects.In “Hepa-tocyte Growth Factor Factor-Scatter Factor and c-Met Eeceptor"" Birkhauser Verlag, 1-29 (1993)

[Publications] K.Matsumoto and T.Nakamura(ed.I.D.Goldbergの分担): "Roles of HGF as a pleiotrpic factor in organ eegeneeration.In“Hepatocyte Growth Factor-Scatter Aactor and c-Met Receptot"" Birkhauser Verlag, PP.225-249 (1993)

[Publications] K.Matsumoto T.Nakamura(eds.Drs.J.B.Griffiths,A.Doyle and D.G.Newellの分担): "Primary and continuous cell culture:Collagenase and Hyaluronidase.In “Protocols in Cell and Tissue Culture"" john Wiley & Sone Limited,England, (1993)

[Publications] T.Nakamura,K.Muzuno,and K.Matsumoto(eds.T.Tsuruta,M.Seno and M.Doyamaの分担): "Hepatocyte growth factor:Mitogen,motogen,morphpgen,and its roles in tissue regeneration In “New Functionalty Materials:Design,Preparation and Control." Elsevier Science Prblishers B.V., (1993)

中村敏一大阪大学, 医学部, 教授 (00049397)