【摘要】 目的研究顺铂作用于A549细胞后,miR-16和bcl-2的表达变化,并探讨两者的相关性。方法采用MTT法测定顺铂对A549细胞的抑制率、锥虫蓝拒染法检测细胞凋亡率,倒置显微镜观察细胞形态,确定合适的药物浓度;然后提取细胞中的miRNA,用实时定量PCR技术定量分析miR-16在对照组细胞和加药组细胞中的表达变化;通过microRNA_org等分析软件预测miR-16的下游调控基因;采用Western blot方法分析细胞中bcl-2蛋白的表达变化。结果在顺铂诱导作用下,A549细胞凋亡率明显增加;miR-16在顺铂作用后的细胞中表达显著升高,而bcl-2蛋白显著低表达。结论顺铂可以促使肺癌A549细胞凋亡;miR-16具有抑癌基因活性,能负调控bcl-2蛋白的表达,参与顺铂致A549细胞死亡的作用。更多还原

【Abstract】 Objective To explore the expression changes of miR-16 and bcl-2 in the A549 lung cancer cell lines treated with Cisplatin.Methods The growth inhibition rate of lung cancer cells was analyzed by MTT assay,the number of dead cells was determined by counting of trypan blue exclusion staining cells and the morphology of A549 cells was observed by the inverted microscope to deter mine the appropriate Cisplatin concentration.Then the miRNA was extracted from the A549 cells,the changes of miR-16 expression in the control group and the dosing group were analyzed by Real-time PCR and miRNA’s downstream regulating genes were evaluated by microRNA——org analysis software.The expression of bcl-2 protein in the two groups was detected by Western blot.Results After Cisplatin treatment,the number of A549 dead cells was significantly increased.The expression of miR-16 in Cisplatin group was significantly higher than that in the control group.However,the expression of bcl-2 protein in Cisplatin group was lower than that in control group.Conclusion Cisplatin induced lung cancer A549 cell apoptosis;miR-16 down-regulated the expression of bcl-2 protein as a tumor suppressor genes,involved in the mechanism of A549 cell death induced by Cisplatin.更多还原