二甲双胍对人肝癌细胞Bel-7402增殖与凋亡的影响

Effects of metformin on proliferation and apoptosis of human hepatocelluar carcinoma cell line Bel-7402

目的:研究二甲双胍抑制人肝癌细胞Bel-7402增殖及调亡的影响,并初步探讨其机制。方法:体外培养人肝癌细胞株Bel-7402,以不同浓度的二甲双胍(10mM、20mM、40mM)干预细胞48h。采用MTT法检测药物对细胞增殖的影响;采用Hoechst 33258荧光染色法和流式细胞术检测细胞凋亡;Real-time PCR检测caspase-3、bax和bcl-2的mRNA水平表达。结果:与对照组相比,不同浓度的二甲双胍作用于Bel-7402细胞48h后,MTT结果显示对细胞增殖有明显抑制作用(P<0.05);Hoechst 33258荧光染色法显示凋亡细胞明显增多;流式细胞术分析显示二甲双胍可明显诱导Bel-7402细胞凋亡,晚期凋亡率分别为3.76%,8.96%和18.67%;caspase-3、bax和bcl-2的mRNA表达水平均上调(P<0.05);以上结果均具有浓度依赖性。结论:二甲双胍能抑制Bel-7402细胞增殖并诱导其凋亡,其机制可能与caspase-3、bax和bcl-2的mRNA表达水平改变有关。

Objective： To investigate the effects of metformin on proliferation and apoptosis in human hepatocelluar carcinoma cell line Bel-7402. Methods： Human hepatocelluar carcinoma cell line Bel-7402 were cultured in vitro and were treated with metformin （10 mM, 20 mM, 40 mM） for 48 h. The growth inhibition rates of Bel-7402 cells were measured by MTT assay. The ceil apoptosis was measured by Hoechst33258 staining and flow cytometry. The expression levels of caspase-3, tax and bcl-2 mRNA were detected by real-time PCR. Results： Metformin （10 mM, 20 mM, 40 mM） markedly deceased the proliferation of Bel-7402 in a dose-dependent manner （P%0. 05 vs. control）. Metformin had obvious apoptosis-inducing activity on Bel-7402 cells. The rates of late apoptotic cells increased to 8. 6 %, 10.5 %, 15.2%, respectively. The expression levels of m spase-3, bax and ＆＇l-2 mRNA were upregulated in a dose-dependent manner （P〈0. 05 vs. control）. Conclusion： Metformin could inhibit the growth and induce apoptosis of Bel-7402 cells mainly via changing expression levels of related gene mRNA.