Experimental Study on Treatment of Cervical Cancer with HPV16-E6-Targeted SiRNA

Objective To study the effects of the HPV16-E6-targeted siRNA on the biological behavior of cervical cancer cells by using RNA interference techniques and the HPV-encoded oncoprotein HPV16-E6 as a target. Moreover,we attempt to elucidate the clinical significance of its experiment. Methods The expression vectors of HPV16-E6-targeted siRNA were constructed and transfected into the HPV16-E6-positive CaSki cells by vitro transfection reagents. The expression levels of the HPV16-E6 mRNA and the mRNA encoded oncoproteins in cervical cancer CaSki cells were detected by RT-PCR and Western blotting,respectively. The expression and activities of the apoptosis-associated molecules were analyzed through cytochrome c assay in order to investigate the molecular mechanism of cell apoptosis induced by HPV16-E6-targeted siRNA. Results RT-PCR assay demonstrated that after the expression vectors of HPV16-E6-targeted siRNA were transiently transfected into the HPV16-E6-positive CaSki cells,a variety of considerable changes occurred:the expression of E6 proteins and mRNA in CaSki cells were down-regulated; Western blotting showed that the expression of anti-apoptotic protein Bcl-2 was also down-regulated; the cytochrome c liberation test displayed its release from mitochondria into cytoplasm,thus inducing the cell apoptosis. Conclusions HPV16-E6-targeted siRNA can significantly inhibit the proliferation of transplanted tumor cells and induce cell apoptosis. It finds out a new approach for the functional study of this important oncoprotein HPV16-E and provides new experimental evidences for the gene-targeted treatment of HPV16-E6-positive tumors,especially exploring a new gene therapy for HPV infection and cervical cancer.