The Infiltrating Development And Apoptotic Molecular Mechanisms Of Prostate Cancer Cells By Silencing NOL3 Gene

Objective: Using Rt2 Profiler PCR Array gene chip technology to detect the difference expression of genes in PCA and BPH tissues.Targeting the NOL3 gene in prostate cancer cells by sh RN A interference technology.And then observed their biological behavior change,and discuss the role of NOL3 gene on apoptosis of prostate cancer cel s.Methods: 1.Collection of PCa and BPH tissues,using Rt2 Profiler PCR Array gene chip technology to detect the difference expression of genes.We verified the larger differences of gene expression by q RT-PCR technology,which had a network of mutual relations.2.We structured the inhibitory lentivirus of NOL3 gene.PL/sh RNA/GFP-homoNOL3-341 transfected prostate cancer cell lines(PC3 and DU145),was transfected;transfected of ester as the negative control group;untreated group for blank group.Observed the transfection efficiency of prostate cancer cell lines by fluorescence microscope.Using q RT-PCR technology to detect the expression of NOL3 gene after transfection;the proliferation of prostate cancer cell lines were detected by MTT assay at 24 h,48h and 72 h after transfection,and the change of tumor cell invasion also was detected by Transwell chamber.The expression of NOL3 proteins in PC3 and DU145 cells were detected by Western bloting;the influence of PC3 and DU145 cell apoptosis after transfection was measured by F low cytometry.Using Caspase 8/9 activity assay kit to detecet the change of caspase 8/9 activity after transfection.Result: 1.Based on Rt2 Profiler PCR Array technology to detect tumor-related genes in PCa and BPH tissue,and with a further verification,we found that AASDHPPT?CASP2?CASP8?CFLAR?CSNK2A1?EHD1?EHD4?NIF3L1?NOL3?SIRPA?TFPT and VASP,a total of 12 genes which had expression difference.2.The transfection efficiency of PL/sh RNA/GFP-homo-NOL3-341 in PC3 and DU145 cells were 90% and 80%,respectively.After transfection,the expression levels of NOL3 gene and NOL3 proteins in PC3 and DU145 cells were significantly lower than the negative control group and blank group(P < 0.05);but the proliferation inhibition rate of PC3 and DU145 cells after transfection ware significantly higher than other two groups(P <0.05),and this effect was in a time-dependent manner.The aggressivity of PC3 and DU145 cells after transfection was obviously lower than negative control group and blank group(P < 0.05).Caspase-8/9 activation level and the apoptosis rate of PC3 and DU145 cells were higher than negative group and blank group(P < 0.05).Conclusion: 1.Rt2 Profiler PC R Array gene chip detection technology can effectively filter out the key genes which associated with occurrence,deve lopment and transfer of prostate cancer.2.The process of occurrence,development and metastasis of prostate cancer is a common regulation by multiple genes and factors,rather than a single factor.3.NOL3 gene can inhibit the apoptosis of tumor cells.Target silencing NOL3 gene and inhibit its expression,can significantly reduce the proliferation and aggressivity of prostate cancer cel s,and also have inhibitory effect on cell apoptosis.