Effects of microRNA -30c knockdown on proliferation and differentiation of P19 cells
10.3760/cma.j.issn.2095-428X.2015.13.009
- VernacularTitle:microRNA-30c 沉默对 P19细胞增殖及分化的影响
- Author:
Xuehua LIU
;
Shasha ZHU
;
Zhangbin YU
;
Chun ZHU
;
Mengmeng LI
;
Shuping HAN
;
Xiaoshan HU
;
Jin'gai ZHU
;
Yuzhu PENG
- Publication Type:Journal Article
- Keywords:
microRNA - 30c;
P19 cell;
Proliferation;
Differentiation
- From:
Chinese Journal of Applied Clinical Pediatrics
2015;(13):992-995
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of microRNA(miRNA)- 30c knockdown on proliferation,diffe-rentiation of P19 cells. Methods miRNA - 30c knockdown plasmid(miRNA - 30c knockdown group)or no - load vector(negative control group)was transfected into P19 cells by lipo2000 and stable cell lines were selected by Blastici-din;Dual luciferase reporter gene system was used to confirm miRNA - 30c knockdown. Cell counting kit - 8(CCK - 8) assay was adopted to detect cell proliferation activity. An inverted microscope was used to observe morphological chan-ges of P19 cell differentiation. Cells were induced to differentiated to myocardiocyte with dimethyl sulfoxide(DMSO). Differentiation marker genes including cTnT,NKX2. 5,GATA4 relative mRNA expression levels were detected with real - time quantitative polymerase chain reaction,respectively. Results Observation of green fluorescent protein ex-pression under a fluorescence microscope indicated similar transfection efficiencies,and miRNA - 30c knockdown re-leased the activity of target gene Gli2. As a result,miRNA - 30c knockdown vector was constructed successfully(P ﹤0. 001). During differentiation of mouse P19 cells into myocardial cells,the beating cell clusters in miRNA - 30c knockdown cells were much lower than those in the control cells,and cTnT,NKX2. 5,GATA4 in miRNA - 30c knock-down cells showed significantly lower expression than those in the control cells( all P ﹤ 0. 05). Conclusions miRNA - 30c inhibits the P19 cell proliferation and differentiation. This study gives us a new insight of heart develop-ment and we need more efforts on exploring the deep function of heart diseases.
