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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 hepatoma cells and its mechanism. Methods Human hepatoma cell line HepG2 was used in the experiment. The effect of HGF on cell proliferation was observed by BrdU cell proliferation enzyme-linked immunosorbent assay. The effect of HGF on S-phase Skp2 expression was detected by Western blot and RT-PCR. The Skp2 eukaryotic recombinant plasmid was transfected into HepG2 cells and stably transfected HepG2 cells overexpressing Skp2 were obtained. The effect of HGF on the proliferation of transfected cells was observed. Results HGF had a concentration-dependent and time-dependent inhibitory effect on the proliferation of HepG2 cells. HGF significantly inhibited the expression of Skp2 protein and mRNA in HepG2 cells. However, HGF did not inhibit proliferation of HepG2 cells overexpressing Skp2. Conclusion HGF has a significant inhibitory effect on HepG2 cells proliferation, and its mechanism is closely related to the down-regulation of Skp2 expression.