Preliminary Study Of Targeting Treatment For Castration Resistant Prostate Cancer Based On BIRC Gene

Objective:To study the expression of BIRC6 gene in prostate cancer and its effect on the biologic behavior of prostate cancer cells in vivo and vitro.To investigate the effects of down-regulation of IAP gene family based on BIRC6 on the biological behavior of prostate cancer cell.Methods: The expression of protein and mRNA of BIRC6 in normal prostate cancer RWPE-1 and prostate cancer cell lines LNCaP,PC-3,C4-2 and DU145 were detected by Western Blot and RT-qPCR.The BIRC6 low expression plasmid was constructed,and the low expression plasmid of BIRC6 was extracted and transfected into PC-3 cells.Western and RT-qPCR were used to detect the expression of BIRC6 protein and mRNA in PC-3 cell line.The cell proliferation of PC-3 with low expression of BIRC6 was detected by MTT and CCK-8.Similarly,the changes of growth,invasion,apoptosis and cell cycle of PC-3 with low expression of BIRC6 were analyzed by monoclonal formation assay,Transwell,Annexin V and flow cytometry,respectively.At the same time,BIRC2 low expression plasmid was designed and transfected/co-transfected with BIRC6 low expression plasmid into PC-3 cells to observe the effects on the above biological behavior.PC-3 cells were transplanted in NOD-SCID mices subcutaneously and the effect of BIRC6-specific antisense oligonucleotides(ASOs)by intraperitoneal injection on tumor growth was observed.Recording and collating experimental data and image,statistical analysis were used with SPSS to analyze the experimental results.Results:The expression of protein and mRNA of BIRC6 in normal prostate cancer RWPE-1 and prostate cancer cell lines LNCaP,PC-3,C4-2 and DU145 were detected by Western Blot and RT-qPCR.We found that the expression of BIRC6 gene in patients with CRPC was significantly increased,so the androgen independent PC-3 cells were chosen as the research object.The BIRC6 low expression plasmid was constructed,and the low expression plasmid of BIRC6 was extracted and transfected into PC-3 cells.Western and RT-qPCR were used to detect the expression of BIRC6 protein and mRNA in PC-3 cell line.The cell proliferation of PC-3 with low expression of BIRC6 was detected by MTT and CCK-8.Similarly,the changes of growth,invasion,apoptosis and cell cycle of PC-3 with low expression of BIRC6 were analyzed by monoclonal formation assay,Transwell,Annexin V and flow cytometry,respectively.At the same time,BIRC2 low expression plasmid was designed and transfected/co-transfected with BIRC6 low expression plasmid into PC-3 cells to observe the effects on the above biological behavior.PC-3 cells were transplanted in NOD-SCID mices subcutaneously and the effect of BIRC6-specific antisense oligonucleotides(ASOs)by intraperitoneal injection on tumor growth was observed.Recording and collating experimental data and image,statistical analysis were used with SPSS to analyze the experimental results.Conclusion: The change of BIRC6 expression can affect the biological behavior of prostate cancer cells,inhibition of BIRC6 combination with BIRC2 can decrease prostate cancer growth in vivo and vitro more effectively.