MicroRNA-34a靶向SIRT1参与阿霉素诱导的心肌细胞凋亡

发布时间：2019-05-28 05:33

【摘要】：目的:研究微小RNA-34a(microRNA-34a,miR-34a)在阿霉素诱导的心肌细胞凋亡中的作用及其作用靶基因。方法:建立阿霉素(doxorubicin,Dox)诱导的大鼠H9c2心肌细胞凋亡模型;TUNEL染色观察H9c2细胞凋亡;双萤光素酶报告实验检测miR-34a与潜在靶基因沉默信息调节因子1(silent information regulator 1,SIRT1)3'端非翻译区(3'-untranslated region,3'UTR)的结合作用;实时荧光定量PCR检测miR-34a和SIRT1 mRNA表达水平,Western blot检测SIRT1和凋亡相关蛋白表达水平。结果:阿霉素处理H9c2细胞之后,细胞发生凋亡,miR-34a的表达显著增强;双萤光素酶报告实验提示miR-34a与SIRT1 3'UTR可相互作用,并证实miR-34a可在转录后水平抑制SIRT1的表达,SIRT1蛋白水平在阿霉素处理的心肌细胞中显著下调;过表达miR-34a及沉默SIRT1均能一致性抑制Bcl-2表达,促进Bax和p66shc的表达,而过表达SIRT1能有效抑制阿霉素诱导的H9c2细胞凋亡。结论:SIRT1是miR-34a的靶基因,并介导了miR-34a在阿霉素诱导的心肌细胞凋亡中的作用。
[Abstract]:Aim: to study the role of microRNA-34a (microRNA-34a,miR-34a) in cardiomyocyte apoptosis induced by doxorubicin and its target gene. Methods: the apoptosis model of H9c2 cardiomyocytes induced by doxorubicin (doxorubicin,Dox) was established, and the apoptosis of H9c2 cells was observed by TUNEL staining. The binding of miR-34a to potential target gene silencing information regulator 1 (silent information regulator 1, 3 'untranslated region (3'-untranslated region,3'UTR) was detected by double luciferase reporting assay. The expression levels of miR-34a and SIRT1 mRNA were detected by real-time fluorescence quantitative PCR., Western blot was used to detect the expression of SIRT1 and apoptosis-related proteins. Results: H9c2 cells were apoptotic and the expression of miR-34a was significantly increased after treatment with doxorubicin. The double luciferase assay suggested that miR-34a could interact with SIRT1 3'UTR, and it was confirmed that miR-34a could inhibit the expression of SIRT1 at post-transcriptional level, and the level of SIRT1 protein was significantly down-regulated in adriamycin-treated cardiomyocytes. Overexpression of miR-34a and silent SIRT1 could inhibit the expression of Bcl-2 and promote the expression of Bax and p66shc, while overexpression of SIRT1 could effectively inhibit the apoptosis of H9c2 cells induced by doxorubicin. Conclusion: SIRT1 is the target gene of miR-34a and mediates the role of miR-34a in adriamycin-induced cardiomyocyte apoptosis.
【作者单位】：南方医科大学;广东省人民医院/广东省医学科学院;
【基金】：国家自然科学基金资助项目(No.81270222;No.81470439;No.91649109) 广东省医学科学研究项目(No.2014A030313635;No.2013B022000083;No.A2015187)
【分类号】：R96

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