Construction of Antisense Human Papillomavirus ( HPV) 18 E6E7 and Green Fluorescent Protein ( EGFP) Eukaryotic Co-expression Vector and Its Expression in HeLa Cervical Carcinoma Cells

Objective 　In order to seek for a therapeutic approach to the diseases caused by HPV or cervix cancer, we const ructed antisense eukaryotic fluorescent expression vector of human papillomavirus ( HPV) 18 E6 E7 and detected their expression in HeLa cervical carcinoma cells. Methods 　HPV18 E6E7 716bp was amplified the by PCR with the full2length HPV-18 cDNA clone vector as the template. Then the PCR product was inserted into eukaryotic fluorescent expressing vector pEGFP in antisense orientation by DNA recombinant technology and further transfected into HeLa cervical carcinoma cells. Expression of HPV18 E6/E7 in t ransfectant s was examined by the fluorescence microscope, RT-PCR and Western blot . Results 　The fluorescence eukaryotic expression vector carrying antisense HPV18 E6 E7 gene fragment was successfully const ructed. At 48h af ter t ransfection of recombinant plasmid, bright green fluorescence was visible and the E6/E7 mRNA and proteins were obviously down regulated in the HeLa cervical carcinoma cells. Conclusion 　Delivery of antisense HPV18 E6 E7 can effectively suppress the expression of E6/E7 oncogenes and is potentially a new approach to t reat HPV infection and cervix cancer.