二甲双胍诱导人胎盘绒毛癌细胞系JAR凋亡的机制初探

The mechanism of metformin induced human choriocarcinoma cell line JAR apoptosis

目的探索二甲双胍诱导人胎盘绒毛癌细胞系JAR凋亡的作用机制。方法将JAR细胞分为对照组和二甲双胍处理组(终浓度5、10、20、40、80mmol/L)处理24h后选取合适的药物浓度。激光共聚焦显微镜检测细胞凋亡,分别采用实时荧光定量聚合酶链反应(Real-time PCR)和免疫蛋白印记(Western blot)检测凋亡相关基因AMPK,p-AMPK,mTOR,Caspase-3,Bcl-2和Bax的变化趋势。结果 40mmol/L的二甲双胍处理能够引发JAR细胞凋亡,并且激活AMPK的磷酸化并下调mTOR的蛋白水平;同时上调Caspase-3和Bax的mRNA及蛋白水平,并且下调Bcl-2的mRNA和蛋白表达,降低Bcl-2和Bax的蛋白水平比例。结论二甲双胍可能是通过AMPK/mTOR和Caspase-3及Bcl-2/Bax两个通路的共同作用诱导JAR细胞发生凋亡。

Objective To investigate the possible mechanism of metformin on apoptosis of human choriocarcinoma cell.Methods The human choriocarcinoma cell line JAR was selected and divided into control and metformin groups(final concentrations were 5,10,20,40 and 80 mmol/L).Confocal microscope was adopted to detect cell apoptosis after 48 h treatment.The mRNA and protein expression of AMPK,p-AMPK,mTOR,Caspase-3,Bcl-2 and Bax were measured by Real-time PCR and Western blot respectively.Results Compared with the control group,apoptosis rate of JAR cells in the metformin group(final concentration was 40 mmol/L)was remarkably increased.Metformin activated AMPK by phosphorylation and inhibited mTOR protein expression,meanwhile mRNA and protein expression levels of Caspase-3 and Bax were significantly increased,but Bcl-2 mRNA and protein expression were significantly decreased.Conclusion Metformin induces the JAR cells to generate apoptosis by the AMPK/mTOR pathways and Caspase-3,Bcl-2/Bax pathways simaltaneously.