【摘要】 目的:探讨miRNA在抗肿瘤药物去甲斑蝥素(DMC)诱导K562细胞凋亡过程中的作用。方法:DMC诱导细胞凋亡后,通过基因芯片技术分析miRNA表达改变,进一步用RT-PCR和定量PCR验证部分miRNA的表达变化;并应用基因信息软件分析miRNA相关的基因,ELISA验证相关基因表达情况。结果:DMC致凋亡组,基因芯片技术检测到290多种miRNA的表达发生明显改变,用RT-PCR和定量PCR分析结果示:在DMC处理组,miR-16、miR-34a和miR-125等表达增高1.5倍以上,而miR-106和miR-150的表达降低60%以上。通过microRNA TargetScan和miRanda软件分析发现癌基因(bcl-2、E2F1、E2F3)和抑癌基因(RB1、p53)表达可能受上述miRNA表达的调控。ELISA结果示:在DMC组,RB1和P53蛋白表达显著增高,而Bcl-2、E2F1和E2F3蛋白的表达显著降低。结论:DMC能诱导K562细胞发生凋亡,并能引起细胞内miRNA及相关基因的表达发生改变。更多还原

【Abstract】 AIM: To explore the role of miRNA expression in the process of antitumor drug demethylcantharidin(DMC)-induced leukemia cell apoptosis. METHODS: K562 cells were treated with DMC to induce apoptosis.Microarray assessment was performed to detect the changes of miRNAs.The expression of miRNAs was further detected by RT-PCR and real-time PCR.The miRNA-relative genes were analyzed by gene information softwares,and their expression was determined by ELISA analysis. RESULTS: The results of microarray showed that more than 290 miRNAs were differentially expressed after DMC treatment.The expression of miR-16,miR-34 and miR-125 increased at more than 1.5 folds in DMC treatment group determined by RT-PCR and real-time PCR analysis,while both miR-106 and miR-150 were down-expressed over 60%.Using microRNA TargetScan and miRanda analysis software,we found that the expression of oncogenes(bcl-2,E2F1,E2F3) and tumor suppressor genes(RB1,p53) may be regulated by the above miRNAs.The expression of RB1 and P53 proteins significantly increased,while Bcl-2,E2F1 and E2F3 proteins were obviously down-regulated after DMC treatment. CONCLUSION: DMC induces K562 cell apoptosis by regulating the expression of miRNAs and their relative genes.更多还原