Inhibitory effects of silencing EZH2 gene expression on cell proliferation of cholangiocarcinoma QBC939cell line and its mechanisms
10.3969/j.issn.1006-5725.2017.10.014
- VernacularTitle:沉默EZH2表达抑制胆管癌QBC939细胞增殖及其机制
- Author:
Shanfeng LIANG
;
Chunyan LI
;
Zhaolan LI
;
Pengjian ZOU
;
Xin GUAN
;
Xiaorong HAN
;
Guansheng HE
- Keywords:
EZH2 gene;
SiRNA;
Cholangiocarcinoma;
Cell proliferation;
Cell senescence
- From:
The Journal of Practical Medicine
2017;33(10):1584-1588
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibitory effects of silencing expression ofEZH2 gene on the cell proliferation of human QBC939 cells and its mechanisms. Methods The targeting siRNA was designed and trans-fected into QBC939cells. The expressions of EZH2 mRNA and protein were detected by real-time qPCR and west-ern blotting,respectively. The ability of cellproliferationwas analyzed by MTT assay and plate clone formation assay. Cell apoptosis and cycle percentage weremeasured by flow cytometry. Cell senescence was assessed byβ-galactosi-dase dyeing.The expressions of H3K27me3,P14ARF,P16INK4a,P53,P21 and E2F1 proteinwere determined by West-ern blotting.Results Compared with the control group ,the expressions of mRNAand protein were significantly elevat-ed in experimental group. The ability of cellproliferation in the experiment group was significantly down regulated , which could also cause a rise of G1/S phase ,but not a marked variation of apoptosis rate. Silencing EZH2 would induce a obvious senescence phenotype in QBC939 cells. EZH2-siRNA transferredcould also down-regulate the expressions of H3K27me3 and E2F1 protein,while up-regulating the expressions of P14ARF,P16INK4a,P53 and P21 protein in QBC939 cells.Conclusions Silencing EZH2 could induce a significant inhibition on cell proliferation of QBC939 cells,the mechanism of which may be associated with the senescence pathway regulation.
