Effect of miR-3619-5p on proliferation of human bladder cancer cell lines EJ and T24
10.3760/cma.j.issn.1000-6702.2017.03.014
- VernacularTitle:微小核糖核酸miR3619-5p对膀胱癌细胞系EJ和T24细胞增殖的影响
- Author:
Senmao LI
;
Jia HU
;
Xiao YU
;
Hua XU
;
Shaogang WANG
;
Zhangqun YE
- Keywords:
MicroRNA;
Bladder Neoplasms;
p21 gene;
RNA activation
- From:
Chinese Journal of Urology
2017;38(3):211-215
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effects of a synthetic miR3619-5p mimics on bladder carcinoma cell lines of EJ and T24 in vitro.Methods EJ and T24 cells were cultured in vitro and treated with three different processing:negative control group(tinfection with dsControl),positive control group(infection with dsP21-322) and the experimental group(infection with miR-3619-5p)during October 2015 to March 2016.Real-time fluorescent quantitative PCR (qPCR) was performed to detect the expression of p21 mRNA,cell cycle protein D1 (CyclinD1) and cell cycle-dependent kinase (CDK4 and CDK6) mRNA.Western Blot method was conducted to evaluate the expression of p21,CyclinD1 and CDK4 and CDK6 proteins;the change of cell cycle was displayed by flow cytometric analysis.Colony formation assay was used to test the ability of single cancer cell clone proliferation.Cell proliferation assay(MTS) was implemented to observed the inhibitive effect of cell proliferative potential.Results qPCR results showed that miR-3619-5p upregulated p21 mRNA expression (P ＜ 0.05),while the expression of CyclinD1,CDK4 and CDK6 were a little lower(P ＜ 0.05) in EJ and T24cells,respectively.Western Blot analysis testified that the expressions of p21,CyclinD1,CDK4 and CDK6 were difference among groups.Flow cytometry displayed that,the G0/ G1 phase increased significantly after transfected with miR-3619-5p and dsP21-322,compared with dsControl group(P ＜ 0.05),indicating that the cell cycle block in G0/G1 phase.Cell colony formation assay certified that the colony formation rates were less in the groups of miR-3619-5p and dsP21-322 than in that of dsControl group(P ＜ 0.05).Cell proliferation assay demonstrated that,cell proliferation ability decreased obviously when transfected with miR-3619-5p and dsP21-322 (P ＜0.05),compared with dsControl group.Conclusions miR-3619-5p could up-regulate the expression of p21 by RNA activation pathway and remarkably induced cell cycle arrest in G0/G1 phase,inhibiting the proliferation of bladder cancer cells.
