Influence Of Retrovirus-mediated HPV16E6-siRNA On The Cervical Cancer Cell Line SiHa

Objective To investigate the effects of retrovirus-mediated siRNA targeting HPV16 E6 gene (HPV16 E6-siRNA) on the HPV 16-positive cervical cancer SiHa cells.Methods SiHa cells were infected with viruses obtained from the retro virus-generating PA317 cells and then were selected with G418 to generate HPV16E6 gene-silencing cell clone SiHa/16E6. The control groups (SiHa/pSUPER group and SiHa group) were designed at the same time. The mRNA levels of HPV16 E6 and E7 were tested by RT-PCR; the protein expression of P53, Rb, caspase 3 and PARP was detected using Western blotting; MTT was performed to study the cellular proliferation activity and their sensitivity to cisplatin.Results Compared with SiHa group, E6 and E7 mRNA expression was suppressed, while p53 and Rb protein expression increased; the cleaved parts of caspase-3 and PARP were both increased; the cellular proliferation activity of SiHa/16E6 40d post-infection decreased; the sensitivity of SiHa/16E6 20d post-infection was higher and the IC50 was calculated as 3.92±2.48μg/ml, whereas there was no significant differences between SiHa/pSUPER group and SiHa group.Conclusion The retrovirus-mediated HPV16 E6-siRNA could effectively silence the expression of E6 and E7 genes, inhibit cell proliferation, induce cell apoptosis and increase cell sensitivity to cisplatin.