小檗碱对肝癌HepG2细胞增殖和凋亡的调节作用

Regulative Effects of Berberine on Proliferation and Apoptosis of Hepatocellular Carcinoma Cell Line HepG2

目的探讨小檗碱抑制肝癌HepG2细胞增殖及诱导细胞凋亡的作用及可能机制。方法活细胞计数(CCK-8)法检测小檗碱对HepG2细胞增殖的抑制作用;流式细胞术检测小檗碱对细胞凋亡及细胞周期的影响;实时荧光定量-聚合酶链反应(RT-PCR)及蛋白印迹法(Western blotting)检测抗增殖基因BTG2及细胞周期蛋白CyclinD1的mRNA和蛋白的表达情况。结果小檗碱可以显著抑制肝癌HepG2细胞的增殖,随着小檗碱作用浓度的升高及作用时间的延长,抑制作用增强(P<0.01);小檗碱可使HepG2细胞周期阻滞在G1期,并促进细胞凋亡,凋亡率与小檗碱的作用时间成正比(P<0.05);随着小檗碱作用时间的延长,抗增殖基因BTG2 mRNA的表达增加(P<0.05);细胞周期蛋白CyclinD1 mRNA的表达则逐渐降低(P<0.01)。结论小檗碱能够抑制肝癌HepG2细胞的增殖,诱导细胞凋亡,其机制可能与其上调抗增殖基因BTG2和下调细胞周期蛋白CyclinD1的表达相关。

Objective To investigate the influence and possible mechanism of berberine on inhibiting proliferation and inducing apoptosis in hepatocellular carcinoma HepG2 cells.Methods The inhibitory effect of berberine on the proliferation of HepG2 cells was detected by cell counting kit-8(CCK-8)method,while the influence of berberine on cell apoptosis and cell cycle were detected by flow cytometry.The expression of protein and mRNA of B cell translocation gene 2(BTG2)and Cyclin D1 were determined by Western blotting and real time fluorescent quantitative PCR(RT-PCR)analysis,respectively.Results The proliferation of HepG2 cells were significantly inhibited by berberine in a dose-and time-dependent manner(P<0.01).Berberine blocked the cell cycle in the G1 phase and promoted cell apoptosis,and the rate of apoptosis was proportional to the duration of berberine action(P<0.05).With the extension of time,the expression of BTG2 mRNA and protein were gradually increased(P<0.05),while the mRNA and protein expression levels of Cyclin D1 were significantly decreased(P<0.01).Conclusion Berberine can inhibit the proliferation and induce apoptosis in HepG2 cells,which may be related to up-regulation of BTG2 expression and down-regulation of Cyclin D1 expression.