MicroRNA-146 a promotes proliferation of rat vascular smooth muscle cells by up-regulating cyclin D1 expression
10.3969/j.issn.1000-484X.2016.07.009
- VernacularTitle:微小RNA-146a上调cyclinD1表达诱导大鼠血管平滑肌细胞的增殖
- Author:
Wei XIONG
;
Yu LUO
;
Shaohong DONG
;
Jianghua LI
;
Bihong LIAO
;
Xinli PANG
;
Linjie LUO
- Publication Type:Journal Article
- Keywords:
Vascular smooth muscle cell;
miRNA-146a;
Gene profile;
p53;
cyclin D1
- From:
Chinese Journal of Immunology
2016;32(7):974-978
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To detect and verifica the gene profile difference of microRNA-146a (miR-146a) and its role in the pro-liferation of vascular smooth muscle cells (VSMCs) by gene chip technology. Methods: Artificially synthesized miR-146a mimics(50 nmol/L) ,miR-146 inhibitor ( 50 nmol/L ) , scramble ( 50 nmol/L ) and PBS were transfected into cultured primary rat VSMCs in vitro. After transfection,Real time PCR was used to measure the levels of miR-146a and the cell counting kit 8(CCK8) was employed to investigate the proliferation of VSMCs. The VSMCs interfered by miR-146a inhibitor or miR-146a control were examined by gene chips and the profile of gene were analyzed by bioinformatics technology to detect the different genes and signal transduction pathway. The changes in mRNAs and proteins were accessed separately by Real time PCR and Western blot. Results: Compared with sham and control VSMCs,miR-146a expression level was significantly decreased in treatment with miR-146a inhibitor(P<0. 01),as well as optical density(OD) was also shown remarkably down regulated simultaneously(P<0. 05). The investigation of gene profile revealed that the p53 signal pathway was up-regulated in VSMCs interfered by miR-146a. The mRNA and protein expression levels of p53, caspase3 and PTEN in p53 signal transduction pathway didn′t show significant differences(P>0. 05),however,the mRNA and protein expression levels of cyclin D1 significantly increased in treatment with miR-146a mimics VSMCs group and decreased in miR-146a inhibitor VSMCs group ( compared with sham VSMCs group, both P<0. 05 ) . Conclusion: Our data indicated that miR-146a may promote the proliferation of rat VSMCs by up-regulating cyclin D1 expression.
