miR—342—3p抑制SM22α启动子转录活性.docVIP

miR—342—3p抑制SM22α启动子转录活性 　[摘要] 目的 探究miR-342-3p是否通过作用于SM22alpha;启动子，从转录水平调控SM22alpha;的表达。 方法 通过软件RNAhybrid分析发现，大鼠SM22alpha;启动子中存在一个miR-342-3p的识别位点。将miR-342-3p mimics与报告基因载体pGL3-SM22alpha;-Promoter共转染293A细胞，通过检测荧光素酶活性来分析miR-342-3p对SM22alpha;启动子转录活性的影响。将miR-342-3p mimics转染血管平滑肌细胞，采用qRT-PCR和Western blot分别检测SM22alpha; mRNA和蛋白质水平，分析miR-342-3p对血管平滑肌细胞中SM22alpha;表达的影响。 结果 与miR-control相比，miR-342-3p能够使SM22alpha;启动子转录活性降低（0.54plusmn;0.03）倍，差异有统计学意义（P lt; 0.05）；与miR-control相比，miR-342-3p能够使血管平滑肌细胞中SM22alpha; mRNA水平降低（0.45plusmn;0.04）倍，SM22alpha;蛋白质水平下降（0.41plusmn;0.05）倍，差异有统计学意义（P lt; 0.05）。 结论 miR-342-3p通过抑制SM22alpha;启动子活性，从转录水平降低SM22alpha;的表达，为miRNA调控血管平滑肌细胞表型转化研究提供了新角度。　　[关键词] microRNA；miR-342-3p；SM22alpha;；启动子　　[中图分类号] R34 [文献标识码] A [文章编号] 1674-4721（2016）08（c）-0012-05　　[Abstract] Objective To explore whether miR-342-3p could repress the expression of SM22alpha; at the transcriptional level by targeting its promoter. Methods There was a potential binding site of miR-342-3p in the SM22alpha; promoter analyzed by using RNAhybrid software. 293A cells were co-transfected with miR-342-3p mimics and pGL3-SM22alpha;-Promoter reporter gene vectors， luciferase activity was detected to analyze whether the SM22alpha; promoter activity is repressed by miR-342-3p. Vascular smooth muscle cells were transfected with miR-342-3p mimics， SM22alpha; mRNA and protein levels were detected by using qRT-PCR and Western blot， to explore the influence of miR-342-3p on the expression of SM22alpha; in vascular smooth muscle cells. Results Compared with miR-control， SM22alpha; promoter transcriptional activity was reduced （0.54plusmn;0.03） folds by miR-342-3p， the difference was statistically significant （P lt; 0.05）. The SM22alpha; mRNA in vascular smooth muscle cells was decreased （0.45plusmn;0.04） folds， and the SM22alpha; protein level was decreased （0.41plusmn;0.05） folds by miR-342-3p， compared with miR-control， the differences were statistically significant （P lt; 0.05）. Conclusion miR-342-3p rep