Effects of adenovirus-mediated interferon-β gene on apoptosis of SMMC-7721 cells

Aim:To study the effects of adenovirus-mediated interferon(AdhIFN-β)gene on apoptosis of SMMC-7721 cells in vitro.Methods:The HEK293A cells were infected with recombinant adenovirus AdhIFN-β and proceed with a large-scale amplification of viral recombinant particles in HEK293A cells.Subsequently the virus was purification and concentrated.The titer of AdhIFN-β was determined by plaque assay in HEK293A cells.SMMC-7721 cells were transduced by AdhIFN-β and AdGFP respectively,and the blank group was used as control.The hIFN-β expression was detected by using immunocytochemical method.The effects of recombinant AdhIFN-β gene inducing SMMC-7721 cells apoptosis was detected by using Hochest 33342 fluorescent staining and flow cytometry.Results:AdhIFN-β was propagated in HEK293A cells and purified using Adeno-XTM virus purification kits.The titers of AdhIFN-β reached 2×1011pfu/mL and more than 95% of SMMC-7721 cells could be infected by 40 multiplicity of infection AdhIFN-β.The results of immunocytochemical method showed obvious hIFN-β protein in SMMC-7721 cells after transfection.The apoptosis of SMMC-7721 cells was obviously observed after transfected with recombinant AdhIFN-β by Hochest 33342 fluorescent staining and flow cytometry compared with that of the control cells,while the apoptosis of other groups cells did not observed obviously.The apoptosis rate of SMMC-7721 cells transduced by AdhIFN-β[(28.27±4.21)%]was higher than that of SMMC-7721 cells transduced by AdGFP[(2.08±0.89)%] and that of SMMC-7721 cells [(1.53±0.70)%](F=377.625,P0.001),while the difference is no significance in statistics science between the apoptosis rate of the later groups.Conclusion:Adenovirus-mediated interferon-beta gene could induce human hepatocellular carcinoma cells apoptosis in vitro.