Hpv 18 E6 Gene-specific Silencing On Hela Cell Proliferation And Apoptosis

OBJECTIVETo investigate the proliferative activity and apoptosis of HeLa cells after HPV18 E6 was knocked down by specific synthetic siRNA.METHODSThe chemically synthetic siRNA targeting HPV18 E6 was transfected into HeLa cells by Lipofectamine 2000, then the mRNA levels of HPV18 E6 and hWAPL (human wings apart-like, hWAPL) were tested by RT-PCR. MTT, immunocytochemistry, flow cytometry and transmission electron microscope were used to detect proliferation, expression of p53 protein, cell cycle & apoptosis and variation of ultramicrostructure, respectively.RESULTSMTT results showed the decrease of proliferative activity in HeLa cells; RT-PCR results suggested lower expression of E6 and hWAPL mRNA compared with control group, which is lowest at 48h after the transfection, while higher level of p53 protein expression was detected by immunocytochemistry; flow cytometry revealed obvious G1 arrest in cell cycle and no significative apoptosis; mitochondria swelling and scarce apoptosis were observed by transmission electron microscope.CONCLUSIONThe chemically synthetic siRNA could effectively silence the expression of E6, lower the level of hWAPL transcription and up-regulate the expression of p53 protein. Finally it induces some changes of HeLa cells, including decrease of proliferative activity, G1 arrest in cell cycle, variation of ultramicrostructure.