国家自然科学基金资助项目(30470784),江苏省功能基因组重点实验室开放基金资助项目(1065)
目的:研究人肝细胞癌中表皮生长因子受体(epidermal growth factor receptor, EGFR)的表达及其细胞内信号转导通路,探讨EGFR在促进肿瘤细胞增殖中的作用。方法:用免疫组化实验检测癌组织中EGFR的表达;用EGF和AG1478处理人肝癌细胞HuH7;用Western blot法检测EGFR、p-EGFR、p-ERK蛋白表达;用WST法检测细胞增长率。结果:17例HCC患者癌组织中,EGFR的阳性表达率为100%;HuH7中EGFR的表达明显高于正常肝细胞HL-7702。外源性EGF(10 μg/L)处理后,ERK和EGFR的磷酸化水平提高,同时细胞生长率提高9.2%,而此作用能被AG1478(20 μmol/L)阻断。结论:人肝细胞癌中,EGFR高表达,EGFR的激活主要通过受体磷酸化完成,活化的p-EGFR可能通过ERK/MAPK下游信号通路传递信息,调控人肝细胞癌的发生发展。
Objective:To investigate the expression of epidermal growth factor receptor(EGFR) and the EGFR signal transduction pathway in human hepatocellular carcinoma(HCC) cells, and to explore the molecular mechanism of EGFR pathway mediating the proliferation of HuH7 cell line. Methods:The expression of EGFR in HCC tissues were evaluated by immunohistochemistry(IHC). HuH7 cells were treated with EGF and AG1478. Western blotting was employed to detect the expressions of EGFR, p-EGFR and p-ERK in HuH7 cells, and the cell viability was determined by the cell proliferation reagent CCK-8. Results:Among 17 cases of HCC tissues,the positive rates of EGFR detected were 100%,while the expression of EGFR in HuH7 was more stonger than that in HL-7702. Exogenous EGF(10 μg/L) enhanced the phosphorylation of ERK and EGFR, and also the growth rate of HuH7 cells at 24 h. But the effect of EGF could be totally interrupted by AG1478(20 μmol/L). Conclusion:Our findings suggested that the expression of EGFR should be positive intensively in human HCC cell line HuH7, and the activation of EGFR depend upon the phosphorylation of EGFR. EGFR-ERK/MAPK signal transduction pathway might play a key role in HuH7 cells, which regulated the development and progression of human HCC cells.
蒋丽,张丽,彭韬,孙秋艳,冷静.表皮生长因子受体信号通路调控人肝细胞癌细胞增殖分子机制的研究[J].南京医科大学学报(自然科学版),2007,(6):523-526