Influences of IFI16 Gene Over-expression on Laryngeal Carcinoma Hep-2 Cells

Abstract: IFI16 gene amplified from RT-PCR was constructed into eukaryotic expression plasmid pVR1012 to obtain pVR1012-IFI16 recombinant plasmid， and then this recombinant plasmid was transfected into Hep-2 cells with Lipofecter transfection. After transfection， the ectopic expression of IFI16 gene in Hep-2 cells was analyzed using semi-quantitative RT-PCR and Western blot methods， Hep-2 cells proliferation influenced by the ectopic expression of IFI16 gene was determined using cell growth curve drawing method and MTT method， and Hep-2 cell cycle and apoptosis influenced by the ectopic expression of IFI16 gene was analyzed using flow cytometry. Semi-quantitative RT-PCR result showed that the mRNA level of IFI16 gene was noticeably increased in pVR1012-IFI16 recombinant plasmid transfected Hep-2 cells， and Western blot result also showed that the protein level of IFI16 gene was obviously increased in pVR1012-IFI16 transfected Hep-2 cells. It was found from cell growth curve assay that Hep-2 cells grew slower than control cells after one day of transfection with pVR1012- IFI16 recombinant plasmid， and the growth rate of Hep-2 cells was obviously slower than control cells when transfected with pVR1012-IFI16 recombinant plasmid for two days and statistically different to that of control cells (P<0.05). MTT result showed that the relative viable cell number of Hep-2 cells was remarkably decreased after 48 h of transfection with pVR1012-IFI16 recombinant plasmid and significantly different to that of control cells (P<0.05 and P<0.01). Flow cytometry analysis disclosed that the ratios of sub-G0 phase cells and apoptotic cells in Hep-2 cells transfected with pVR1012-IFI16 recombinant plasmid for 48 h were obviously increased comparing to that of control cells. The above results illustrated that the pVR1012-IFI16 recombinant plasmid could ectopicly express IFI16 gene in Hep-2 cells and the ectopicly expressed IFI16 gene could inhibit Hep-2 cells proliferation， retard Hep-2 cells cycle in sub G0 and induce Hep-2 cells apoptosis.

CSTR: 32200.14.cjcb.2012.02.0006