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Roles of microRNA-29a in expression of Bcl-2 and Mcl-1 in rat cardiomyocytes
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF PATHOPHYSIOLOGY
- Issue:
- 11
- DOI:
- 10.3969/j.issn.1000-4718.2012.11.002
- Key Word:
- MicroRNA-29a;细胞凋亡;B细胞白血病/淋巴瘤-2;髓样细胞白血病-1
Abstract： 目的:探讨microRNA-29a(miR-29a)对大鼠心肌细胞(CM细胞)B细胞白血病/淋巴瘤-2(Bcl-2) 和髓样细胞白血病-1(Mcl-1)表达的调控机制.方法:体外培养新生Sprague-Dawley大鼠CM细胞和人胚肾细胞株293T.合成大鼠miR-29a的拟似物(mimic)和抑制剂(inhibitor).用脂质体Lipofectamine RNAiMAX转染miR-29a mimic进入CM细胞,转染 48 h后分别用实时荧光定量PCR和Western blotting检测Bcl-2和Mcl-1 mRNA和蛋白的表达变化.构建Bcl-2 和Mcl-1 的萤光素酶报告基因载体(DNA质粒),转染293T 细胞(DNA质粒和miRNA共转染)和CM细胞(miRNA和 DNA 质粒先后转染),双萤光素酶报告基因系统检测萤光素酶的表达变化.结果:CM细胞转染miR-29a mimic 48 h后,Bcl-2 和Mcl-1 mRNA和蛋白的水平均下调 (P<0.05).双萤光素酶报告基因系统显示,在293T细胞中,miR-29a可特异抑制带有 bcl-2和mcl-1 3'UTR上野生型识别元件的报告基因表达(P<0.05),在CM细胞中,抑制内源性的miR-29a水平能促进bcl-2和mcl-1 3'UTR上野生型识别元件的报告基因表达.结论:抗凋亡基因bcl-2和mcl-1是miR-29a的靶基因.miR-29a可能通过作用于Bcl-2和Mcl-1实现对心肌细胞凋亡的调控作用,具体效应仍待进一步阐明.
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