Hepatocyte growth factor promotes migration of human hepatocellular carcinoma via phosphatidylinositol 3-kinase
Hepatocyte growth factor promotes migration of human hepatocellular carcinoma via phosphatidylinositol 3-kinase
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DOI：
10.1023/a:1006685218766
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发表时间：
1999-01-01
影响因子：
4
通讯作者：
Okamoto, E
中科院分区：
文献类型：
--
作者：
Nakanishi, K;Fujimoto, J;Okamoto, E
肝细胞生长因子（HGF）已知是上皮细胞的一种强效有丝分裂原和促动原。肝细胞癌（HCC）经常发生转移，且HCC细胞高度表达c - Met/HGF受体。本研究的目的是通过c - Met探究与HGF对HCC细胞的促动效应相关的信号通路。使用HCC细胞系（Hep3B、HepG2、PLC和Huh - 7）以及从患者获取的HCC细胞进行趋化活性的博伊登小室试验以及免疫沉淀和免疫印迹研究。HGF以剂量依赖的方式刺激Hep3B、HepG2和Huh - 7细胞的运动性，同时伴有c - Met的酪氨酸磷酸化以及磷脂酰肌醇3 - 激酶（PI3 - K）的激活。一种酪氨酸激酶抑制剂（染料木黄酮）和一种PI3 - K抑制剂（渥曼青霉素）阻止了HCC细胞的迁移。然而，蛋白激酶C（PKC）抑制剂卡弗斯汀C（它是磷脂酶Cγ（PLCγ）的下游靶点）并不能阻止迁移。HGF还刺激了从3名患者获取的HCC细胞的迁移，而渥曼青霉素阻止了这些细胞的迁移。这些结果表明，HGF通过激活PI3 - K使c - Met发生酪氨酸磷酸化，从而刺激HCC细胞的迁移。
Hepatocyte growth factor (HGF) is known to be a potent mitogen and motogen for epithelial cells. Hepatocellular carcinoma (HCC) often metastasizes, and the c-Met/HGF receptor is highly expressed by HCC cells. The aim of this study was to investigate the signaling pathways associated with the motogenic effect of HGF on HCC cells via c-Met. HCC cell lines (Hep3B, HepG2, PLC, and Huh-7) and HCC cells harvested from patients were used for the Boyden chamber assay of chemotactic activity as well as for immunoprecipitation and immunoblotting studies. HGF stimulated the motility of Hep3B, HepG2, and Huh-7 cells in a dose-dependent manner in association with tyrosine phosphorylation of c-Met and activation of phosphatidylinositol 3-kinase (PI3-K). A tyrosine kinase inhibitor (genistein) and a PI3-K inhibitor (wortmannin) prevented the migration of HCC cells. However, migration was not prevented by calphostin C, an inhibitor of protein kinase C (PKC), which is a downstream target of phospholipase C gamma (PLC gamma). HGF also stimulated the migration of HCC cells obtained from three patients, while wortmannin prevented the migration of these cells. These results indicate that HGF stimulates the migration of HCC cells through the tyrosine phosphorylation of c-Met via activation of PI3-K.