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Hepatitis C virus F protein inhibits cell apoptosis by activation of intracellular NF‐κB pathway
Author(s) -
Shao Shengwen,
Wu Wenbin,
Bian Zhongqi,
Yu Jianguo,
Zhao Ping,
Zhao Lanjuan,
Zhu Shiying,
Qi Zhongtian
Publication year - 2009
Publication title -
hepatology research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.123
H-Index - 75
eISSN - 1872-034X
pISSN - 1386-6346
DOI - 10.1111/j.1872-034x.2008.00452.x
Subject(s) - apoptosis , microbiology and biotechnology , flow cytometry , transfection , luciferase , hepatitis c virus , intracellular , plasmid , tumor necrosis factor alpha , blot , chemistry , biology , virus , gene , virology , immunology , biochemistry
Aim: To observe the influence of HCV F protein on apoptosis of HepG2 cells, and explore the association between F protein and NF‐κB signal pathway. Methods: HCV 1b F gene containing HepG2‐F cells and HCV 1b C gene containing HepG2‐C cells were treated with 100 IU/mL TNF‐α, 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. Results: (i) With the treatment of TNF‐α 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‐α 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‐α 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‐κB‐luc, and then treated with TNF‐α (100 IU/mL) for 18 h, showed a pcDNA3.1‐F dose‐dependent increase. Conclusion: HCV F protein can over‐activate NF‐κB signal pathway, which makes HepG2‐F cells able to resist TNF‐α induced apoptosis.