Joint Study on the Tumorigenicity of HGF and c-met Receptor

HGF与c-met受体致瘤性的联合研究

• 批准号: 05044174
• 负责人: MIZUNO Kensaku
• 金额: $ 6.4万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05044174/
• 关键词: Hepatocyte Growth Factor; HGF; c-Met; Oncogene; HGF Converting Enzyme; Tyrosine Kinase; LIM Motif; Receptor; 転移; 人工変異体; プロテアーゼ阻害剤

Hepatocyte growth factor (HGF) is a pleiotropic factor, which act as a mitogen, motogen, and morphogen for various epithelial cells. The c-Met protooncogene product was identified as the receptor for HGF.In this study, we analyzed the tumorigenicity of HGF and c-Met receptor. Various sarcoma cell lines were found to express high level of c-Met receptor, indicating that the HGF-Met autocrine signaling contributes to the tumorigenic process of sarcoma.HGF converting enzyme, which converts an inactive single-chain precursor (pro-HGF) to a two-chain active form HGF,was purified and identified as a heterodimer protein composed of a 65-kDa heavy chain and a 32-kDa light chain. A synthetic peptide derivative, Ac-Lys-Gln-Leu-Arg-chloromethylketone, containing the sequence of pro-HGF cleavage site strongly inhibited the converting enzyme activity, and the mitogenic activity of pro-HGF.However, this inhibitor could not block the growth of sarcoma cells, probably because of the instability in the conditioned medium. Using the chicken c-sea tyrosine kinase gene (the gene closely related to c-Met) as a probe, we obtained the cDNAs encoding three novel protein kinases, c-Sea, Sky and LIMK.c-Sea is highly related to the c-Met receptor and seems to be a receptor for a HGF-related growth factor. Sky is a novel receptor tyrosine kinase related to Axl/Ufo/Ark. Identification of the ligands for c-Sea and Sky is the subject of ongoing studies. LIMK is a cytoplasmic serine/threonine kinase containing two repeats of LIM/double zinc finger motif at the N-terminus and a protein kinase domain at the C-terminus. LIMK may be involved in the oncogenic process through interactions with other LIM-containing proteins.

肝细胞生长因子（HGF）是一种多效性因子，可作为多种上皮细胞的有丝分裂原、促生长因子和形态发生素。 c-Met原癌基因产物被鉴定为HGF的受体。在本研究中，我们分析了HGF和c-Met受体的致瘤性。各种肉瘤细胞系被发现表达高水平的c-Met受体，表明HGF-Met自分泌信号传导有助于肉瘤的致瘤过程。HGF转换酶，它将无活性的单链前体（pro-HGF）转化为一种双链活性形式 HGF，经纯化并鉴定为由 65 kDa 重链和 32 kDa 轻链组成的异二聚体蛋白。含有pro-HGF裂解位点序列的合成肽衍生物Ac-Lys-Gln-Leu-Arg-氯甲基酮强烈抑制转换酶活性以及pro-HGF的有丝分裂活性。然而，该抑制剂不能阻断肉瘤细胞的生长，可能是因为条件培养基的不稳定。以鸡c-sea酪氨酸激酶基因（与c-Met密切相关的基因）为探针，获得了编码c-Sea、Sky和LIMK三种新型蛋白激酶的cDNA。c-Sea与c-Met高度相关。 -Met 受体，似乎是 HGF 相关生长因子的受体。 Sky 是一种与 Axl/Ufo/Ark 相关的新型受体酪氨酸激酶。 c-Sea 和 Sky 配体的鉴定是正在进行的研究的主题。 LIMK 是一种细胞质丝氨酸/苏氨酸激酶，在 N 末端含有两个 LIM/双锌指基序重复，在 C 末端含有一个蛋白激酶结构域。 LIMK 可能通过与其他含有 LIM 的蛋白质相互作用参与致癌过程。

项目成果

M.Lyon et al.: "The interaction of HGF with heparan sulfate.Elucidation of the major heparan sulfate structural determinants." J.Biol.Chem.269(in press). (1994)

M.Lyon 等人：“HGF 与硫酸乙酰肝素的相互作用。硫酸乙酰肝素主要结构决定因素的阐明。”

K.Mizuno et al.: "Hairpin-loop and second kringle domain are essential for heparin-binding and biological activity of hepatocyte growth factor." J.Biol.Chem.269. 1131-1136 (1994)

K.Mizuno 等人：“发夹环和第二个三环结构域对于肝素结合和肝细胞生长因子的生物活性至关重要。”

K.Mizuno et al.: "Purification and characterization of HGF-converting enzyme:activation of pro-HGF." Biochem.Biophys.Res.Commun.198. 1161-1169 (1994)

K.Mizuno 等人：“HGF 转换酶的纯化和表征：HGF 前体的激活。”

K.Ohashi et al.: "Cloning of the cDNA for a novel receptor tyrosine kinase,Sky,predominantly expressed in brain." Oncogene. 9. 699-705 (1994)

K.Ohashi 等人：“克隆主要在大脑中表达的新型受体酪氨酸激酶 Sky 的 cDNA。”

T.Nakamura et al.: "New Functionality Materials,Volume B:Synthesis and Function Control of Biofunctionality Materials." Elsevier Science Publishers, pp.313-320 (1993)

T.Nakamura 等人：“新功能材料，B 卷：生物功能材料的合成和功能控制”。