Effects of E2F-1 gene silencing on cisplatin chemosensitivity of human gastric cancer SGC-7901/DDP cells
10.3969/j.issn.1673-4254.2013.12.04
- VernacularTitle:E2F-1基因沉默可增强人胃癌耐药细胞株SGC-7901/DDP对顺铂的敏感性
- Author:
Chao LIAN
1
;
Jie YANG
;
Xiaotong WANG
;
Yubo XIE
;
Qiang XIAO
Author Information
1. 广西医科大学第一附属医院胃肠腺体外科
- Keywords:
E2F-1;
gastric cancer;
cisplatin;
chemosensitivity;
survivin;
Bcl-2
- From:
Journal of Southern Medical University
2013;(12):1727-1732
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of E2F-1 gene silencing on the chemosensitivity of human gastric cancer SGC-7901/DDP cells to cisplatin and explore the underlying mechanism. Methods Gastric cancer SGC-7901/DDP cells were transfected with the recombinant lentivirirus vector Lv-shRNA-E2F-1 for E2F-1 gene silencing, with cells transfected with the control recombinant lentivirirus vector Lv- shRNA- NC as the negative control. MTT assay was used to evaluate cisplatinchemosensitivity of the cells, and the cell apoptosis rate and cell cycle distribution were detected by flow cytometry. The mRNA and protein expressions of E2F-1 and apoptosis-related genes (survivin and Bcl-2) were detected by RT-PCR and Western blotting. Results MTT assay showed that the IC50 of cisplatinwas significantly lowered in Lv-shRNA-E2F-1-transfected cells compared with the negative and blank control cells (P<0.05). Lv-shRNA-E2F-1 transfection caused significant cell cycle arrest in G0/G1 phase and induced obvious cell apoptosis. Compared with Lv-shRNA-NC group and the blank control group, Lv-shRNA-E2F-1 group showed significantly lowered expressions of E2F-1 mRNA by 45.0%and 41.3%and E2F-1 protein by 66.7%and 70.5%, survivin mRNA by 30.3%and 28.7%and survivin protein by 56.5%and 53.6%, and Bcl-2 mRNA by 76.6%and 76.8% and Bcl-2 protein by 74.6% and 79.9%, respectively. No significant difference was found in the measurements between Lv-shRNA-NC group and the blank control group (P>0.05). Conclusion E2F-1 gene silencing can enhance cisplatinchemosensitivity of gastric cancer SGC-7901/DDP cells possibly by down-regulating survivin and Bcl-2 expressions, suggesting the value of E2F-1 as a new chemotherapeutic target for gastric cancer.
