Hepatitis C virus F protein inhibits cell apoptosis by activation of intracellular NF‐κB pathway

Abstract

To observe the influence of HCV F protein on apoptosis of HepG2 cells, and explore the association between F protein and NF-kappaB signal pathway. HCV 1b F gene containing HepG2-F cells and HCV 1b C gene containing HepG2-C cells were treated with 100 IU/mL TNF-alpha, and analyzed by flow cytometry, Western blotting, and dual luciferase reporter assay. Empty plasmid pcDNA3.1(+) containing HepG2-3.1 cells were used as control. (i) With the treatment of TNF-alpha for 18 h, the apoptosis rates (AR) of HepG2-F and HepG2-3.1 cells were 0.41% (+/- 0.11%) and 37.43% (+/- 2.03%) respectively, while that of HepG2-C was 4.07% (+/- 0.18%). At 36 h after TNF-alpha treatment, the AR of HepG2-F and HepG2-3.1 cells were 10.03% (+/- 0.41%) and 44.63% (+/- 3.37%), and that of HepG2-C was 14.95% (+/- 0.85%). (ii) After the treatment of TNF-alpha for 0.5-18 h, the p65 contents in the whole cells of HepG2-F and HepG2-3.1 showed no significant difference (P = 0.34, t = 1.08), while the p65 contents in the nucleus of HepG2-F and HepG2-3.1 cells were 3.8-1.9 times and 1.8-1.0 times higher than that in the non-treated cells (P = 0.013, t = 4.25). (iii) The relative luciferase unit (RLU) of the HepG2 cells, co-transfected with pcDNA3.1-F and pNF-kappaB-luc, and then treated with TNF-alpha (100 IU/mL) for 18 h, showed a pcDNA3.1-F dose-dependent increase. HCV F protein can over-activate NF-kappaB signal pathway, which makes HepG2-F cells able to resist TNF-alpha induced apoptosis.