Effect of miR-21 over-expression on proliferation, migration and diffe-rentiation of c-Kit+ cardiac stem cells
10.3969/j.issn.1000-4718.2018.06.001
- VernacularTitle:过表达微小RNA-21对c-Kit+心脏干细胞增殖、迁移及分化的影响
- Author:
Yan WANG
1
;
Bei SHI
;
Wen-Wen DENG
;
Dong-Mei WANG
;
Jin SHENG
;
Wen-Ming CHEN
Author Information
1. 遵义医学院附属医院心内科
- Keywords:
MicroRNA-21;
Cardiac stem cells;
Cell proliferation;
Cell migration;
Cell differentiation
- From:
Chinese Journal of Pathophysiology
2018;34(6):961-968
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To explore the effect of microRNA (miR)-21 on proliferation, migration and differentiation abilities of c-Kit+ cardiac stem cells (CSCs). METHODS:c-Kit+ CSCs were cultured and selected by the methods of en-zyme digestion and magnetic bead separation. miR-21 mimics (50 nmol/L) and mimics negative control ( MNC) were transfected into c-Kit+CSCs with Lipofectamine?2000. The cells was divided into 3 groups:control group:c-Kit+ CSCs without any pretreatment; MNC group:the cells were transfected with MNC for 48 h; mimics group:the cells were trans-fected with miR-21 mimics for 48 h. qPCR was used to assess the expression of miR-21 in each group. CCK-8 and EdU as-says were used to determine the cell proliferation. qPCR and immunofluorescence were used to detect the differentiation in each group. Scratch assay was adopted to explore the migration ability of the cells. RESULTS:The expression of c-Kit in the c-Kit+ CSCs were 90.8%, with 0.6% of CD45 and 0.5% of CD34. A significant increase in miR-21 expression was observed when the cells were transfected with miR-21 mimics for 48 h ( P<0.05). CCK-8 and EdU assays showed that miR-21 significantly increased cell proliferation as compared with MNC group and control group (P<0.05). No difference in the expression of Nkx2.5, CD31 and α-SMA at mRNA and protein levels was observed, and no difference of the migra-tion ability in 3 groups of the c-Kit+ CSCs was found. CONCLUSION:Over-expression of miR-21 significantly promotes the proliferation of c-Kit+ CSCs, without any effect on the cell migration and differentiation.
