Objective To investigate the impact of express of Cancer/Testis Antigen Family 45 Member A1(CT45A1)on the in vitro proliferative,migratory and invasive capacities of prostate cancer cells,and to dissect its functional mechanism.Methods The levels of CT45A1 expression in human prostate cancer cells with varying invasive capacity were measured using real-time fluorescent quantitative polymerase chain reaction and Western blot analyses.Lentiviral transduction was used to generate prostate cancer cells with CT45A1 knockdown.The effect of CT45A1 knockdown on proliferation,migration,and invasion was assessed using the MTT method,wound healing assay,and Transwell assay,respectively.The levels of E-cadherin and vimentin were measured as markers of epithelial-mesenchymal transition(EMT),along with two transcription factors that regulate EMT,Twist and Zeb-1.The measurements were taken using real-time fluorescent quantitative polymerase chain reaction and WB analysis.Results The results of fluorescence quantitative PCR and WB experiments indicate that the relative expression of CT45A1 mRNA(4.37±0.34)and the protein expression level of CT45A1 in PC-3 cells of human prostate cancer were significantly higher than that in LNCap cells.Additionally,the CT45A1 mRNA and protein expression levels in shCT45A1#1 and shCT45A1#2 cells were lower than those in shScram cells,and the differences were all statistically significant(P<0.05).After silencing the CT45A1 gene,the proliferation level of PC-3 cells did not change significantly(P>0.05).However,the migration and invasion abilities were decreased,and the expression of Vimentin and Zeb-1 were also decreased,while the expression of E-cadherin was up-regulated.All of these differences were statistically significant(P<0.05).Conclusions CT45A1 may augment the motility and invasiveness of prostate cancer PC-3 cells in vitro by triggering EMT mediated by Zeb-1.
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