三羟异黄酮对肝癌细胞HepG2的ADAM17表达的调节

Genistein's regulation to the gene ADAM17 expression on hepatocarcinoma cell line HepG2

目的:观察三羟异黄酮(genistein)对肝癌细胞HepG2的生长增殖及ADAM17基因表达的影响。方法:培养肝癌细胞HepG2,加入三羟异黄酮特异性阻断酪氨酸蛋白激酶(TPK),MTT法观察genistein对HepG2细胞生长增殖的影响,PCR分析ADAM17基因水平变化,免疫组化法观察细胞水平ADAM17蛋白表达的变化。结果:经过genistein处理的肝癌细胞HepG2生长增殖受到明显抑制,在48、72和96小时有统计学意义(P<0.01);而且ADAM17的基因表达和蛋白表达都受到明显抑制(P<0.05),抑制程度呈现时间依赖性关系。结论:三羟异黄酮可以通过抑制TPK通路抑制肝癌细胞HepG2的生长增殖,并且抑制ADAM17基因和蛋白表达水平,从而间接调节HepG2细胞生长信号的传递。

Objective：To explore the effects on the proliferation and gene ADAM17 expression of hepatocareinoma cell line HepG2 after treated with genistein. Methods： Culture HepG2 cells in vitro and treat the cells with genistein to block tyrosine protein kinase （TPK）, then assay the proliferation by MTT, analyse the expression on gene level by reverse transcription PCR and on protein level by cellular immunohistochemistry at 24, 48, 72 and 96 hours following treatment. Results： After the HepG2 cells were treated with genistein, proliferation was significantly inhibited compared to the control groups at 48,72 and 96 hours （P 〈0.01 ）, and the expression difference of gene ADAM17 of the cells was statistically inhibited both in gene level and protein level （P 〈0. 05）, and further, the inhibited degree showed time - dependent. Conclusion： Genistein can inhibit the proliferation of HepG2 ceils and lower the gene ADAM17 expression by inhibiting the TPK pathway, thereby indirectly regulate the growth signal transmission of hepatocarcinoma cells.