Influence of hepatitis B virus X gene on apoptosis of hepatic cells mediated by Fas
10.3760/cma.j.issn.1007-3418.2017.06.007
- VernacularTitle: 乙型肝炎病毒X基因对Fas介导的肝细胞凋亡的影响
- Author:
Hewen WU
1
;
Kuan LI
;
Yanli ZENG
;
Yi KANG
;
Junping LIU
;
Huibin NING
;
Jia SHANG
Author Information
1. Department of Infectious Disease, Henan Provincial People’s Hospital, Zhengzhou 450003, China
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus X gene;
HepG2 cells;
Hepatocytes;
Apoptosis;
Fas
- From:
Chinese Journal of Hepatology
2017;25(6):424-428
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the influence of hepatitis B virus X gene (HBx) on apoptosis of hepatic cells mediated by Fas in HePG2 cells.
Methods:HBx eukaryotic vector pcDNA3.1(+)-X was transfected into HEPG2 cells with lipofectamine, and the null vector pcDNA3.1(+) and untransfected HEPG2 were used as normal controls. The cells were collected 72 h after transfection, and the expression of HBx mRNA and protein was determined using RT-PCR and Western blot, respectively. The mRNA expression of apoptosis-related genes Bcl-2 and Bax mRNA was also determined using RT-PCR. Cytotoxicity and apoptosis were evaluated using CCK-8 and flow cytometry, respectively, after HepG2-HBx and HepG2-3.1 cells were treated with stimulatory monoclonal antibody anti-Fas CH11. The t test was used for pairwise comparison.
Results:The cell line HepG2-HBx was successfully established, as confirmed by RT-PCR and Western blot, and RT-PCR results showed that HepG2-HBx cells had significantly higher expression of Bcl-2 mRNA than HepG2-3.1 and HepG2 cells (P < 0.05), but had significantly lower expression of Bax mRNA than HepG2-3.1 and HepG2 cells (P < 0.05); CCK-8 and flow cytometry showed that anti-Fas CH11 had a lower cytotoxicity to HepG2-HBx cells and allowed for a lower apoptosis rate of HepG2-HBx cells compared with HepG2-3.1 and HepG2 cells.
Conclusions:HBx can inhibit apoptosis of hepatic cells mediated by the Fas pathway.
