基础医学与临床 ›› 2016, Vol. 36 ›› Issue (11): 1537-1541.

• 研究论文 • 上一篇    下一篇

miR-31及其靶基因LATS2在大鼠肥大心肌细胞中的表达变化

曾俊义,张婉,丁露,魏云峰,郑泽琪,文通,易达松   

  1. 南昌大学第一附属医院
  • 收稿日期:2016-04-21 修回日期:2016-07-01 出版日期:2016-11-05 发布日期:2016-10-24
  • 通讯作者: 魏云峰 E-mail:wyf5629@163.com
  • 基金资助:
    江西省卫生计生委科技计划

Expression of miR -31 and its target gene LATS2 in rat hypertrophic cardiomyocytes

  • Received:2016-04-21 Revised:2016-07-01 Online:2016-11-05 Published:2016-10-24

摘要: 目的 观察大鼠心肌细胞肥大过程中miR-31及其靶基因LATS2的表达变化。方法 构建双萤光素酶基因报告系统鉴定miR-3l对LATS2的靶向作用。体外培养原代大鼠心肌细胞,给予10-6mol/L血管紧张素II(AngII)刺激构建体外心肌细胞肥大模型,RT-qPCR检测miR-31、LATS2及心肌肥大基因ANP与β-MHC表达,心肌细胞F肌动蛋白荧光探针染色观察心肌细胞形态变化,Western blot进一步检测LATS2蛋白表达。结果 miR-3l可与LATS2-3' UTR基因片段特异性结合并使荧光素酶活性显著降低。10-6mol/LAngII干预48 h后可检测到心肌细胞肥大基因ANP及β-MHC表达上调(P<0.01和P<0.05),干预96 h后心肌细胞表面积明显增大。伴随心肌细胞的肥大变化,miR-31表达显著上调(P<0.01),LATS2在基因及蛋白水平均明显下调(P<0.05)。结论 伴随大鼠心肌细胞的肥大变化,miR-31表达显著上调,而LATS2在基因及蛋白水平均明显下调。

关键词: miR-31, LATS2, 心肌细胞, 肥大

Abstract: ObjectiveTo observe the expression of miR-31 and its target gene LATS2 in the hypertrophy process of rat cardiomyocytes.MethodsTargeting regulation of miR-31 on LATS2 was identified by a dual luciferase gene reporter system. Rat primary cardiomyocytes were isolated and cultured in vitro, and hypertrophic model of cardiomyocytes was constructed by administering 10-6mol/L AngII. MiR-31,LATS2 and hypertrophy genes ANP,β-MHCwere detected by RT-qPCR.Morphology of the cardiomyocyteswas observed by fluorescence staining to F-actin.LATS2 protein was further detected by western blot.ResultsMiR-3l could specifically bind to 3'UTR of LATS2,which made the luciferase activity decrease significantly.Hypertrophy genes ANP and β-MHCwere up-regulated at 48 hour and the area of cardiomyocytes was increased significantly at 96 hour after administration of 10-6mol/LAngII. Accompanied with the hypertrophic changes of cardiomyocytes, miR-31 was significantly up-regulated, while LATS2 in gene and protein levels were reduced obviously.ConclusionsAccompanied with the hypertrophic changes of rat cardiomyocytes, miR-31 was significantly up-regulated, while LATS2 in gene and protein levels were reduced obviously.

Key words: miR - 31, LATS2, cardiomyocytes, hypertrophy

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