Effects Of Human Papillomavirus 16 E6 Small Interference RNA On Cervical Carcinoma Cells SiHa

Objective To establish the human papillomavirus (HPV) 16 oncogene E6 silencing cellclones and investigate the biological characteristics of the cervical carcinoma cell line SiHacells after E6 gene was stably silenced by HPV 16 E6 small interference RNA (HPV 16 E6siRNA).Methods The specific siRNA of HPV 16 E6 was synthesized and transfected into SiHa cellsby lipofectamine. The cell proliferative activity, HPV 16 E6 messenger RNA (mRNA)expression and the changes of cell cycle and apoptosis were measured before and after thetransfection by proliferation assay, reverse transcriptase-polymerase chain reaction and flowcytometry, respectively.Resμlt The SiHa cells that stably silenced E6 oncogene were obtained. Compared withnon-infected groups, the expression of E6 mRNA level was reduced remarkably afterpassages 2, 15 and 25 respectively (P＜0.01). The apoptosic rates were significantly increasedfrom 1.8% (non-infected groups) to 11.3 %(passage 2),7.7% (passage 15) and 5.7% (passage25) (P＜0.01). The percentages of G1 phase cells significantly increased in clone cells afterpassages 2, 15 and 25 (P＜0.01). On the contrary, the percentages of S phase was decreased,compared with non-infected group cells (P＜0.01). The proliferation was suppressedsignificantly in clone cells.Conclusion We successfμlly established the cell clones in which the HPV16+E6 gene wasstably silenced. The resμlts demonstrate that sequence-specific siRNA had inhibitory effecton cell growth, mRNA expression and inductive effects on cell apoptosis. Our study indicatesthat siRNA HPV16 E6 can be used as a novel therapeutic approach for cervical cancer.