miR3423p抑制SM22α启动子的转录活性.docVIP

miR3423p抑制SM22α启动子的转录活性 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：miR3423p抑制SM22α启动子的转录活性 1 1 资料与方法 3 293A细胞转染与报告基因分析 4 2 结果 5 3 讨论 6 文2：非小细胞肺癌中hMLH1启动子的甲基化 8 1资料与方法 8 参考文摘引言： 12 原创性声明（模板） 14 文章致谢（模板） 14 正文 miR3423p抑制SM22α启动子的转录活性 文1：miR3423p抑制SM22α启动子的转录活性 [Abstract] Objective To explore whether miR-342-3p could repress the expression of SM22α at the tracriptional level by targeting its promoter. Methods There was a potential binding site of miR-342-3p in the SM22α promoter analyzed by using RNAhybrid software. 293A cells were co-trafected with miR-342-3p mimics and pGL3-SM22α-Promoter reporter gene vecto， luciferase activity was detected to analyze whether the SM22α promoter activity is repressed by miR-342-3p. Vascular smooth muscle cells were trafected with miR-342-3p mimics， SM22α 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 SM22α in vascular smooth muscle cells. Results Compared with miR-control， SM22α promoter tracriptional activity was reduced （±） folds by miR-342-3p， the difference was statistically significant （P ）. The SM22α mRNA in vascular smooth muscle cells was decreased （±） folds， and the SM22α protein level was decreased （±） folds by miR-342-3p， compared with miR-control， the differences were statistically significant （P ）. Conclusion miR-342-3p represses the expression of SM22α at the tracriptional level by targeting its promoter， which provides a novel pepective for the study of miRNA in vascular smooth muscle cell phenotypic switch regulation。 [Key words] microRNA； miR-342-3p； SM22α； Promoter microRNA（miRNA）是一类内源性的、长约22个核苷酸的调控性非编码RNA。miRNA的主要作用模式是通过碱基不完全互补配对的方式介导RNA诱导沉默复合体（RNA-induced silencing complex，RISC）与胞质中靶mRNA的3UTR结合，从转录后水平抑制靶基因的翻译[1]。值得关注的是，成熟miRNA不但分布在细胞质中，还广泛分布在细胞核内，并且很多miRNA在核内的浓度远高于在胞质中的浓度[2-3]。研究证实，胞核中的miRNA通过与基因启动子结合，从转录水平调控靶基因的表达。例如，miR-320通过与POLR3D基因启动子结合，抑制其转录[4]；miR-373与E-cadherin启动子结合在转录水平诱导基因的表达[5]；miR-744和miR