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Chinese Journal of Pathology ; (12): 822-825, 2008.
Article in Chinese | WPRIM | ID: wpr-315061

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of RNA interference (RNAi) targeting E6AP on the proliferation and apoptosis of HeLa cells.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>SiRNA targeting can effectively suppress the expression levels of E6AP mRNA, corresponding with a proliferation inhibition and an enhanced apoptosis of HeLa cells.</p>


Subject(s)
Female , Humans , Cell Line, Tumor , Cell Proliferation , Gene Expression Regulation, Neoplastic , Genetics , Gene Silencing , HeLa Cells , RNA Interference , RNA, Small Interfering , Genetics , Pharmacology , Reverse Transcriptase Polymerase Chain Reaction , Ubiquitin-Protein Ligases , Metabolism , Uterine Cervical Neoplasms , Genetics , Metabolism
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