[SiRNA inhibition of E6AP expression in cervical cancer cells].
摘要
OBJECTIVE: To study the effect of RNA interference (RNAi) targeting E6AP on the proliferation and apoptosis of HeLa cells. METHODS: HeLa cells were cultured and divided into 3 groups: blank control group, cells transfected with nonsense siRNA (small interference RNA), and cells transfected with specific E6AP siRNA. The expressions of E6AP mRNA and protein were detected by RT-PCR and Western blot before and after the transfection respectively. Cell proliferation was determined by methylthiazolyl tetrazolium (MTT). The cell apoptosis index was assessed by flow cytometry. RESULTS: Upon treatment with E6AP siRNA for 24, 48 and 72 h, the expression level of E6AP mRNA decreased 33%, 72% and 70% than siRNA treated group. The protein expression levels in 48 h and 72 h E6AP siRNA groups decreased 38%, 59% comparing with those of the nonsense siRNA treated group (P < 0.05). The proliferative capacity of cells transfectd with E6AP siRNA was significantly lower than blank control group (F = 101.38, P < 0.05) and siRNA treated group (F = 38.64, P < 0.05). The apoptosis index of HeLa cells treated with E6AP siRNA was significantly higher than that of the nonsense siRNA (F = 41.48, P < 0.05) and the blank control group (F = 86.36, P < 0.05). CONCLUSION: SiRNA targeting can effectively suppress the expression levels of E6AP mRNA, corresponding with a proliferation inhibition and an enhanced apoptosis of HeLa cells.