Effects Of Apoptosis And Cell Proliferation By SiRNA Targeting Against HIF-2α Gene On Human Prostate Cancer PC-3Cell Under Hypoxia

Objective：1. Putting cobalt chloride into culture medium to modeling chemistry hypoxia toinvestigate the expression of HIF-2α(Hypoxia-inducible factor-2alpha) in human prostatecancer PC-3cell under hypoxia．2. To investigate the effects of apoptosis and cell proliferation by siRNA targeting againstHIF-2α (Hypoxia-inducible factor-2alpha) gene on the expression in human prostate cancerPC-3cell under hypoxia.Methods:1.The effects of HIF-2α Gene on the proliferation of PC-3cell were estimated by MTT.Semi quantitative reverse transcription-poly merase chain reaction(RT-PCR) was performed todetect the expression of HIF-2α mRNA in human prostate cancer PC-3cell exposed during thephase of hypoxia Induced by CoCl2，the relations of the quantity-efficiency and thetime-efficiency were analyzed．All that in order to elect the best time and concentrate to CoCl2modeling chemistry hypoxia.2. Human prostate carcinoma cell PC-3hypoxia induced by CoCl2．Tests were made ofcontrol groups，CoCl2groups，HIF-2α siRNA+CoCl2groups, empty liposome+CoCl2groups.Semi quantitative RT-PCR was performed to detect the expression of HIF-2α mRNA inhuman prostate carcinoma cell PC-3when it is48hours．The protein expression level ofHIF-2α was measured by Western Blotting when it is48hours．RT-PCR was performed todetect the expression of HIF-2α mRNA at the same time．The effects of apoptosis and cellproliferation were detected by FCM and MTT assay.Results：1. The best model of hypoxia was induced by100μmol/L CoCl2when it is48hours． 2. There were expressions of HIF-2α in human prostate carcinoma cell PC-3. The mRNAexpression level of HIF-2α and the protein expression level of HIF-2α under hypoxia inducedby CoCl2were higher than that of untreated by CoCl2(P<0.05). The levels of mRNA andprotein of HIF-2α were suppressed significantly after treated with HIF-2α siRNA(P<0.05)．There was not statistically significant between CoCl2groups and empty liposome+CoCl2groups; The proliferation potential and apoptosis in human prostate carcinoma cell PC-3was not statistically significant between control groups and CoCl2groups (P<0.05),but theproliferation potential and apoptosis were changed significantly after transfecting HIF-2αsiRNA(P>0.05), and there was not statistically significant between CoCl2groups and emptyliposome+CoCl2groups．Conclusion：1. HIF-2α can over-express in human prostate carcinoma cell PC-3inducing by CoCl2.2. HIF-2α siRNA can effectively reduce the HIF-2α mRNA expressions of the humanprostate carcinoma cell PC-3,and make corresponding protein expression reduced. The PC-3survival rate was droped significantly, and appearing cells apoptosis after transfecting HIF-2αsiRNA. show that HIF-2α can inhibit the apoptosis of human prostate carcinoma cell PC-3．...