RNA interference silencing EZH2 gene strengths the sensitivity of human hepatic multidrug-resistant cancer cells to 5-Fu chemotherapy
10.3760/cma.j.issn.1007-631X.2012.08.015
- VernacularTitle:RNA干扰沉默EZH2基因增强人肝癌多药耐药Bel/Fu细胞对氟尿嘧啶的敏感性
- Author:
Yi ZHANG
;
Bo TANG
;
Rui LIANG
;
Guangpu LIU
;
Chen LIU
;
Liming WANG
- Publication Type:Journal Article
- Keywords:
Liver neoplasms;
Gene silencing;
RNA interference;
Multidrug resistance-associated proteins
- From:
Chinese Journal of General Surgery
2012;27(8):660-663
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo study the impact of EZH2 silence on the sensitivity of human hepatic multidrug-resistant cancer cells Bel/Fu to 5-Fu.MethodsBel/Fu cells were cultured in vitro; EZH2 siRNA was used to interfere EZH2 expression; RT-PCR and Western blot was used to detect the efficiency of interference.MTT assay was used to detect the cellular growth inhibitory rate; Annexin V-FITC/PI double staining was used to detect the apoptosis rate of cells ; Flow cytometry was to analyze cell cycle ; Western blot analysis was used to detect the expression of multidrug resistance-associated protein MDR1 after silencing EZH2.The experiment set up four groups:control group,5-Fu treatment group,EZH2 siRNA treatment group,5-Fu combined with EZH2 siRNA treatment group. ResultsThe expression of EZH2 was greatly decreased after 24 h in the combined group,the apoptotic inhibitory rate by MTT was 43.17％ ± 3.81％,higher than other three groups; the apoptotic rate in the combined group by Flow cytometry was 30.4％ ± 1.77％,markedly higher than other three groups.The cell cycle of the combined group detected by Flow cytometry was 69.16％ ±2.31％ of cells in the combined group at G1 phase,the percentage was higher than other three group,30.76％ ± 1.29％ at S,G2 and M phases,lower than other three groups,indicating the cell cycle was blocked at G1 phase.MDR1 protein level in the combined group was lower than other groups.ConclusionsSilencing EZH2 strengths the sensitivity of Bel/Fu cells to 5-Fu,probably by a mechanism decreasing the expression of MDR1.
