Effects of human papillomavirus 16 E6-specific short hairpin RNA on cervical carcinoma cell

Objective To Find a most effective short hairpin RNA(shRNA) for human papillomavirus(HPV) 16 E6 mRNA and investigate the effects of the HPV16E6 shRNA on cervical carcinoma cell.Methods HPV16 E6-specific hairpin siRNA expression vectors were constructed,sequenced and transfected into the cervical carcinoma SiHa cells,stable expression cell populations were selected with G418,pooled and passaged.HPV16 E6 mRNA levels in siRNA transfected cells were determined by Real-time RT-PCR and the most effective siRNA was screened.p53,p21 proteins were examined by western blot.Results HPV16 E6A siRNA was selected as the most effective siRNA,which could provide a 90% reduction of HPV16 E6 mRNA in the transfected cells.HPV16 E6A hairpin siRNA expression vector could effectively inhibit the expression of HPV16 E6 mRNA at 4 months after transfection.p53,p21 proteins levels increased greatly in transfected cells.The E6A siRNA transfected cells showed a proliferation inhibition.Conclusions HPV16 hairpin siRNA expression vector can stable express siRNA and effectively inhibit HPV viral genes expression in SiHa cells for more than 4 months after transfection.The E6A siRNA can inhibit cell proliferation.