顺铂对LTEP-a-2细胞中miR-34b、Bcl-2、Smad3表达的影响

Effect of cisplatin on MiR-34b,Bcl-2 and Smad3 in LTEP-a-2 cells

目的探讨顺铂对LTEP-a-2细胞内miR-34b、Bcl-2、Smad3表达的影响,明确顺铂的抗癌机制。方法体外培养LTEP-a-2细胞,实验组加入不同浓度顺铂,阴性对照组为不加顺铂的细胞;倒置显微镜观察各组细胞形态,台盼蓝染色法计算细胞存活率,MTT法测定顺铂对LTEP-a-2细胞的抑制率;实时定量PCR检测miR-34b;western blot方法分析细胞中Bcl-2及Smad3蛋白的表达变化。结果随着顺铂作用剂量的增加,LTEP-a-2细胞凋亡增加,miR-34b在顺铂作用后的细胞中的表达呈上升趋势,Bcl-2及Smad3的表达显著下降。结论顺铂可能通过上调miR-34b的表达,进一步抑制Bcl-2及Smad3的表达,导致LTEP-a-2细胞的凋亡。

Objective To explore the effect of Cisplatin on the expression of miR-34 b,Bcl-2 and Smad3 in LTEP-a-2 cells,and explore the antitumor mechanism of cisplatin.Methods LTEP-a-2 cells were cultured in vitro.The experiment groups were treated with different concentrations of Cisplatin,while the contrast groups without Cisplatin;cells were observed under one microscope,cell survival rates were calculated by blue staining method and cell growth inhibition rates were analyzed by MTT assay;then miR-34 bwas amplifed by Real-time PCR and the expressions of Bcl-2 and Smad3 were detected by western blot.Results After treatment of Cisplatin,apoptotic LTEP-a-2 cells and miR-34 bexpression were upregulated,while Bcl-2and Smad3 expressions were downregulated.Conclusions Cisplatin might inhibit the expression of Bcl-2 and Smad3,and induce LTEP-a-2 cells apoptosis by upregulating miR-34 b.