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Objective To observe the effects of HCV protein,NS3 and NS5A on IFN-βin HepG2 cells and its regulation mechanism.Methods Human liver hepatocellular carcinoma cells HepG2 were transfected with recombinant eukaryotic plasmid pcDNA3.1/myc-His-core,NS3 or NS5A to overexpress these proteins,and the expression of IFN-βwere detected by qRT-PCR,Western blotting and ELISA.Luc2P reporter plasmids pGL4.10-IFNβ-P were constructed and transfected into HepG2 cells,and the activity of IFN-βpromoter were determined through luciferase assay for regulation mechanism study.Results Both mRNA level and protein expression of IFN-βwere significantly decreased(P<0.05)in the presence of NS3 or NS5A protein.Luciferase assay revealed that NS3 or NS5A protein downregulated IFN-βpromoter activity(P<0.05).Meanwhile,HCV core protein had little effect on IFN-βexpression.Conclusions HCV protein NS3 and NS5A could inhibit innate IFN-βexpression and thus escape immune selection and hinder the host immune responses.
Objective To observe the effects of HCV protein, NS3 and NS5A on IFN-β in HepG2 cells and its regulation mechanism. Methods Human liver hepatocellular carcinoma cells HepG2 were transfected with recombinant eukaryotic plasmid pcDNA3.1 / myc-His-core, NS3 or NS5A to overexpress these proteins, and the expression of IFN-βwere detected by qRT-PCR, Western blotting and ELISA. Lucp2P reporter plasmids pGL4.10-IFNβ-P were constructed and transfected into HepG2 cells, and the activity of IFN-βpromoter were determined through Results Both mRNA level and protein expression of IFN-β predomotor activity (P <0.05) in the presence of NS3 or NS5A protein. Luciferase assay revealed that NS3 or NS5A protein downregulated IFN- 0.05). Meanwhile, HCV core protein had little effect on IFN-βexpression.Conclusions HCV protein NS3 and NS5A could inhibit innate IFN-βexpression and thus escape immune selection and hinder the host immune responses es.