miR-31及其靶基因LATS2在大鼠肥大心肌细胞中的表达  

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

作　　者：曾俊义[1] 张婉[1] 丁露[1] 魏云峰[1,2] 郑泽琪[1,2] 文通[2] 易达松 

机构地区：[1]南昌大学第一附属医院江西省高血压病研究所,江西南昌330006 [2]南昌大学第一附属医院心血管内科,江西南昌330006

出　　处：《基础医学与临床》2016年第11期1537-1541,共5页Basic and Clinical Medicine

基　　金：江西省卫生计生委科技计划(20161019)

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。方法构建双荧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-q PCR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/L AngⅡ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。Objective To observe the expression of miR-31 and its target gene LATS2 in the hypertrophy process of rat cardiomyocytes. Methods Targeting 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-MHC were detected by RT-q PCR. Morphology of the cardiomyocytes was observed by fluorescence staining to F-actin. LATS2 protein was further detected by Western blot. Results miR-31 specifically bound to 3＇UTR of LATS2,which decreased luciferase activity significantly. Hypertrophy genes ANP and β-MHC were up-regulated at 48 hour and the area of cardiomyocytes was increased significantly 96 hours after administration of 10- 6mol / L AngⅡ. Accompanied with the hypertrophic changes of cardiomyocytes,miR-31 is significantly up-regulated,while LATS2 in gene and protein levels are reduced obviously. Conclusions Accompanied with the hypertrophic changes of rat cardiomyocytes,miR-31 is significantly up-regulated,while LATS2 of gene and protein level reduces obviously.

关 键 词：miR-31 LATS2 心肌细胞 肥大 

分 类 号：R542.2[医药卫生—心血管疾病]