HPV18 E6E7 antisense fluorescence eukaryotic expression vector and its expression in cervical cancer HeLa cells.docVIP

 PAGE \* MERGEFORMAT 16 HPV18 E6E7 antisense fluorescence eukaryotic expression vector and its expression in cervical cancer HeLa cells Author: Secretary Marnie, Wang Wei, Tian Xun, Luo Ai-month, LI Chun-xiao, Wang Juan, LU Yun-Ping, Shi-Xuan Wang, Martin 【Abstract】 Objective To construct the HPV18-type E6E7 antisense fluorescence eukaryotic expression vector, and to observe their cervical cancer HeLa cells, HPV18 E6 and E7 gene expression, to explore the clinical anti-sense technology for the treatment of the possibility of HPV infection and cervical cancer . Methods The HPV18-based whole-genome plasmid as a template, PCR amplified HPV18-type E6E7 area 716bp fragment, using recombinant technology to insert a fluorescent reverse fragment of eukaryotic expression vector pEGFP  C1, EcoR I digestion and sequencing; Lipid transfer method using the recombinant plasmid pEGFP  HPV18E6E7as (EGFP  18AS) transfected HeLa cervical cancer cell lines by RT  PCR and western blot detection of cell E6, E7 mRNA and protein expression. The results successfully constructed HPV18 E6E7 antisense fluorescence eukaryotic expression vector EGFP  18AS, by lipofection HeLa cells, 48h after the inverted fluorescence microscope, clearly visible under the green fluorescence, and the cell E6, E7 mRNA and protein expression were significantly down-regulated. Conclusion fluorescent antisense eukaryotic expression vector can effectively inhibit HPV18 E6, E7 oncogene expression, for the treatment of HPV infection and cervical cancer provides a new approach. Keywords: human papillomavirus type 18; E6/E7; Antisense; EGFP; cervical cancer Cervical cancer is a common malignancy of women in the global context, its incidence second only to breast cancer, but its incidence in many developing countries ranked first, accounting for 25% of the total number of female malignancies [1] . Studies have shown that 99% of cervical cancer can be detected in human papilloma vi