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，对多种上皮细胞具有促分裂、促运动和促形态发生的作用。c-Met原癌基因产物被鉴定为HGF的受体，本研究分析了HGF与c-Met受体的致瘤性。多种肉瘤细胞系均表达高水平的c-Met受体，表明HGF-Met自分泌信号参与了肉瘤的发生过程。HGF转化酶是一种由65 kDa重链和32 kDa轻链组成的异源二聚体蛋白，可将无活性的单链前体（pro-HGF）转化为双链活性形式的HGF。含促肝细胞生长素原（pro-HGF）切割位点序列的合成肽衍生物Ac-Lys-Gln-Leu-Arg-chloromethylketone对促肝细胞生长素原（pro-HGF）的转化酶活性和促有丝分裂活性有明显的抑制作用，但该抑制剂不能抑制肉瘤细胞的生长，可能与其在条件培养基中的不稳定性有关。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三种新蛋白激酶的cDNA。Sky是一种与Axl/Ufo/方舟相关的新型受体酪氨酸激酶。鉴定c-Sea和Sky的配体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/双锌指基序重复序列，在C-末端含有一个蛋白激酶结构域。LIMK可能通过与其他含LIMK的蛋白相互作用参与致癌过程。

项目成果

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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.Mizuno,et al.: "Comprehensive Endocrinology,The Pituitary Gland,2nd.ed." Raven Press, 520 (1995)

K.Mizuno 等人：“综合内分泌学，垂体，第二版”。

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。”