Effects Of ShRNA Inhibition Of Activation Of Cytidine Deaminase (AID) On The Phenotype Of Prostate Cancer Cell Line C4-2

Objective: To investigate the expression of activation-induced cytidine deaminase(AID)in normol prostate,benign prostatic hyperplasia and prostate cancer tissue samples,and the expression of prostate cancer cell C4-2.The effect and mechanism of AID on biological behaviors such as invasion,migration,proliferation and apoptosis of prostate cancer cell C4-2.Methods: Immunohistochemical staining and Western-blot method were used to detect the expression of AID in normal prostate,benign prostatic hyperplasia and prostate cancer tissue samples and prostate cancer cell C4-2.The expression of E-cadherin was detected by immunofluorescence technique.Using gene interference technology to inhibit AID expression in prostate cancer cell line C4-2,screening for the best inhibitory monoclonal cell line.Western-blot method was used to detect the expression of AID in C4-2 cells and blank control group(C4-2 group)and negative control group(NC group,C4-2 cells transfected with empty vector sequence)after sh RNA inhibition of AID expression.Methyltransferase inhibitor(5-AZA-d C)stimulates Wnt/β-catenin signaling pathway and EMT-related protein expression in a monoclonal cell group(Monoclonal 6 group,a monoclonal cell strain that inhibits the expression of AID).The proliferation of Monoclonal6 cells in C4-2 cells was detected by plate cloning method.The apoptosis rate of Monoclonal6 cells in C4-2 cells was detected by flow cytometry.The invasion and migration ability of Monoclonal6 cells were detected by transwell chamber assay.Results: The expression of AID in prostate tissue and prostate cancer cell C4-2 was high.The expression of AID in normal prostate and benign prostatic hyperplasia tissue was very low,the difference was statistically significant(*P<0.05),and the expression level of E-cadherin in the above three human tissue samples was opposite to that of AID;the expression of AID in Monoclonal6 group was significantly lower than that in C4-2 or NC group after inhibition of AID expression(*P<0.05),and the expression levels of GPR177,β-catenin in the Wnt/β-catenin pathway and N-cadherin and vimentin in EMT were significantly decreased(*P<0.05),and MMP14 and SDF1 also significantly decreased their expression due to inhibition of AID(*P<0.05);the invasiveness and migration ability of Monoclonal group 6 after inhibition of AID expression was significantly lower than that of C4-2 group or NC group(*P<0.05),and the proliferative capacity was also significantly reduced(*P<0.05),but the apoptotic rate increased significantly(*P<0.05).The expression of GPR177,β-catenin and N-cadherin was up to the pre-inhibition level after 5-AZA-d C stimulation of Monoclonal6 cells.Conclusions:AID is highly expressed in human prostate cancer tissues and C4-2 cells,the abnormally high expression of AID in human prostate cancer cell line C4-2 can increase its proliferation ability and inhibit its apoptosis,AID may mediate Wnt/β-catenin signaling pathway to regulate EMT-induced invasion and migration of prostate cancer cells through demethylation.