myocardin通过激活miR-93-5p促进人主动脉血管平滑肌细胞分化  

Myocardin Promoting Differentiation of Human Aortic Vascular Smooth Muscle Cells by Activating miR-93-5p

作　　者：史丹阳 刘美玲 刘翔[1] 张建 张同存[1] 王楠[1] SHI Danyang;LIU Meiling;LIU Xiang;ZHANG Jian;ZHANG Tongcun;WANG Nan(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,Tianjin Key Laboratory of IndustrialMicrobiology,College of Biotechnology,Tianjin University of Science & Technology,Tianjin 300457,China)

机构地区：[1]工业发酵微生物教育部重点实验室天津市工业微生物重点实验室天津科技大学生物工程学院,天津300457

出　　处：《天津科技大学学报》2019年第2期19-24,共6页Journal of Tianjin University of Science & Technology

基　　金：天津市自然科学基金资助项目(17JCZDJC33600);天津市高等学校创新团队培养计划资助项目(TD13-5015)

摘　　要：myocardin是促进平滑肌细胞分化的重要转录因子.微小RNA(microRNAs,miRNA)在近年来已经证实参与多种生物学过程,包括平滑肌的表型转化.然而在平滑肌分化过程中myocardin与哪些microRNA具有相关性仍有待于阐明.通过将外源性myocardin转入人主动脉血管平滑肌细胞(human aortal vascular smooth muscle cells,HA-VSMCs)中,利用免疫印迹(Western blot)检测分化标志基因肌动蛋白α2(actin alpha2,ACTA2)和肌动蛋白相关蛋白(smooth muscle 22 alpha,SM22α)的表达;利用实时荧光定量PCR(Real-timePCR)检测过表达或敲低myocardin对miR-93-5p水平的影响;构建miR-93-5p启动子的荧光素酶报告质粒,利用荧光素酶报告分析证实myocardin对mi R-93-5p启动子的转录激活作用.上述结果证实myocardin可能通过激活miR-93-5p的转录,上调其表达,进而促进血管平滑肌细胞的分化.Myocardin is an important transcription factor, which promotes the differentiation of vascular smooth muscle cells. It has been reported that microRNAs are involved in multiple biological processes, including smooth muscle cell phe-notypic switching. However, what microRNAs are associated with myocardin during smooth muscle cell differetiation process is still unclear. In this research, exogenous myocardin was transfected into human aortal vascular smooth muscle cells(HA-VSMCs), and then the expression of contractive genes actin alpha 2(ACTA2)and smooth muscle 22 alpha (SM22 α)were detected by Western blot. By using fucntion gain or loss approach, the effect of myocardin on the level of miR-93-5p was investigated and its level was tested with Real-time PCR. By constructing luciferase reporter plasmid containing promoter miR-93-5p, the effect of myocardin on the activity of miR-93-5p promoter was detected by luciferase assay. These data showed that myocardin promoted the differentiation of VSMCs by activating the transcription of miR-93-5p and upregulating its level.

关 键 词：MYOCARDIN 人主动脉血管平滑肌细胞 表型转化 miR-93-5p 

分 类 号：Q28[生物学—细胞生物学]