Construction of three lentiviral vectors expressing HPV16 E5, E6 or E7 oncogene and respective infection of oral epithelial cells with the vectors / 安徽医科大学学报

ABSTRACT

Objective To provide the basis for the study of HPV16 E5 gene inthe pathogenesis of oral squamous cell carcinoma by constructing three lentiviral vectors expressing HPV 16 E5, E6 or E7 oncogene and transfecting o-ral epithelial cells with the vectors.Methods HPV16 E5, E6 or E7 oncogene was amplified using PCR and liga -ted into the lentiviral vector pLVX-AcGFP-N1 respectively to construct vectors pLVX -AcGFP-E5, pLVX-AcGFP-E6 and pLVX-AcGFP-E7, then respectively cotransfected 293T cells with packaging plasmids , viral supernatant was collected to respectively transfect oral epithelial cells .Afterpuromycin screening,oral epithelial cells with HPV 16 E5, E6,or E7 oncogene transfection were constructed , then reverse transcription PCR and western blotassays were performed for verifying HPV16 E5, E6 or E7 expression.Results pLVX-AcGFP-E5, pLVX-AcGFP-E6 and pLVX-AcGFP-E7 were successfully constructed, oral epithelial cells expressing HPV 16 E5, E6 or E7 oncogene wereacquired, HPV16 E5,E6 or E7 expression was confirmed in oral epithelial cellsthrough reverse transcription PCR and western blot assays.Conclusion Three lentiviral vectors expressing HPV16 E5, E6 or E7 oncogene can successfully infect oral epithelial cells .