miRNA在去甲斑蝥素致K562细胞凋亡中的作用研究  

Effect of miRNAs on apoptotic leukemia cells induced by demethylcantharidin

作　　者：张帅[1] 李有杰[1] 张超[1] 岳真[1] 张文娟 刘在贵[1] 谢书阳[1] 

机构地区：[1]滨州医学院生物化学教研室,医学分子遗传学研究所,山东烟台264003 [2]烟台山医院,山东烟台264003

出　　处：《中国病理生理杂志》2011年第3期499-503,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30801324);山东省科技厅基金资助项目(No.ZR2009CQ033No.2007BS03048);滨州医学院学术骨干及科研基金资助项目(No.BY2008KJ17)

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。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.

关 键 词：去甲斑蝥素 细胞凋亡 微小RNA 基因表达 

分 类 号：R733.7[医药卫生—肿瘤]