HGF下调Skp2表达抑制人肝癌细胞增殖的实验研究  

The effect of HGF-mediated Skp2 down-regulation on suppression of proliferation of human hepatocellular carcinoma cells

作　　者：贺亮[1] 刘冰阳[2] 丁宏[1] 丁奎[1] 张浩[1] 

机构地区：[1]中国医科大学附属第一医院普通外科,辽宁沈阳110001 [2]中国医科大学93期临床医学七年制,辽宁沈阳110001

出　　处：《中国普通外科杂志》2009年第7期699-702,共4页China Journal of General Surgery

基　　金：国家自然科学基金资助项目(30572114);高等学校博士学科点专项科研基金(20070159001);教育部留学回国人员科研启动基金(2005383);辽宁省教育厅高等学校科学技术研究项目(05L509);辽宁省博士启动基金资助项目(20051040)

摘　　要：目的探讨肝细胞生长因子(HGF)对人肝癌细胞增殖的作用及其机制。方法人肝癌细胞株HepG2用于实验。通过BrdU细胞增殖酶联免疫实验观察HGF对细胞增殖的作用,采用Western印迹和RT-PCR法检测HGF对S期激酶结合蛋白2(Skp2)表达的影响。将Skp2真核细胞重组表达质粒转染HepG2细胞并获得稳定转染的过表达Skp2的HepG2细胞,观察HGF对该转染细胞增殖的影响。结果HGF对HepG2细胞的增殖具有浓度依赖性和时间依赖性的抑制作用;HGF明显抑制HepG2细胞表达Skp2蛋白和mRNA。但HGF对过表达Skp2的HepG2细胞无增殖抑制作用。结论HGF对HepG2细胞具有明显的增殖抑制作用,其作用机制与下调Skp2表达密切相关。Objective To investigate the effect of hepatocyte growth factor （ HGF ） on the proliferation of human hepatocellualr carcinoma cells, and the mechanism of HGF-induced proliferation inhibition. Methods Human hepatocellualr carcinoma cell line HepG2 were treated with different concentrations of HGF for different time periods, and the proliferation of these cells was examined by colorimetric BrdU cell proliferation enzyme-linked immunosorbent assay. The expression of S-phase kinase associated protein 2 （ Skp2 ） was examined using Western blot and RT-PCR. Plasmids pcDNASkp2 was introduced into HepG2 cells, then the clones showing up-regulation of Skp2 were selected, and the effect of HGF on the proliferation in these clones was investigated. Results HGF inhibited the proliferation of hepatoma cells in a dose and time dependent manner. The expression of Skp2 was significantly suppressed by HGF. Furthermore, HGF did not suppress the proliferation of HepG2 cells transfected with Skp2. Conclusions This study suggests that HGF could inhibit HepG2 cell proliferation, and the down-regulation of Skp2 could be closely related to this suppressed proliferation.

关 键 词：癌 肝细胞 肝细胞生长因子 SKP2 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤]