| Objective:Prostate cancer(PCa)is a common malignant tumor of the urinary system.It is currently second only to lung cancer as an important cause of morbidity and mortality in men.In this study,The Cancer Genome Atlas(TCGA)database analysis showed that suppressor APC domain containing 2(SAPCD2)gene was differentially expressed between prostate cancer tissues and normal tissues,but the expression and role of SAPCD2 in prostate cancer have not been reported.Therefore,this study is to investigate the expression of SAPCD2 in prostate cancer tissues and normal prostate tissues and its correlation with clinicopathological data.Secondly,the effect of SAPCD2 gene expression on the biological function of prostate cancer DU145 and PC-3 cells was investigated.Methods:1.Screening of genes related to the prognosis of prostate cancer through TCGA database;the expression of selected target genes was analyzed by gene expression profiling interactive analysis 2(GEPIA2)online database.2.The expression of SAPCD2 protein in tissue microarray(including 57 cases of prostate cancer tissues,20 cases of benign prostatic hyperplasia tissues and 2 cases of normal prostate tissues)was detected by immunohistochemistry,and the correlation between SAPCD2 protein expression and age,distant metastasis,T stage,lymph node metastasis,and Gleason grade of prostate cancer patients was analyzed.3.The background expression of SAPCD2 RNA in DU145,PC-3 and RWPE-1 cells was detected by q PCR.4.The prostate cancer PC-3 and DU145 cells were transfected with lentiviruses containing three different RNA interference sequences,and q PCR was used to screen the most effective interference series of SAPCD2 gene.5.The selected RNA interference series were transfected into DU145 and PC-3 cells,and the transfection efficiency was observed by fluorescence microscope.The transfection efficiency of DU145 and PC-3 cells was verified by q PCR and WB experiments at RNA and protein levels,respectively.6.The effect of silencing SAPCD2 gene on the proliferation of prostate cancer DU145 and PC-3 cells was detected by CCK-8 assay;7.The effects of silencing SAPCD2 gene on the migration and invasion of prostate cancer DU145 and PC-3 cells in vitro were detected by Transwell migration assay and Transwell invasion assay;8.The effect of silencing SAPCD2 gene on apoptosis of prostate cancer DU145 and PC-3 cells was detected by flow cytometry.Results:1.The resμlts of TCGA database analysis showed that SAPCD2 was an up-regulated differential gene in prostate cancer;GEPIA2 online database analysis showed that the expression of SAPCD2 in prostate cancer tissues was higher than that in adjacent tissues(P < 0.05),and the prognosis of patients in the high expression group was worse than that in the low expression group(P < 0.05).2.The expression of SAPCD2 protein in tissue microarray detected by IHC showed that the expression of SAPCD2 in prostate cancer was higher than that in normal prostate tissue(P < 0.05).3.The expression of SAPCD2 m RNA in RWPE-1 cells was significantly lower than that in PC-3 and DU-145 cells(P < 0.001).4.The results showed that compared with the control group,the gene knockout rates of sh SAPCD2-2 and sh SAPCD2-3 in the transfection group were the highest.By comparing the resμlts of three groups of RNA interference sequences,it was found that sh SAPCD2-2 gene knockout rate was the best in DU145 and PC-3,so sh SAPCD2-2 was used in subsequent experiments.5.WB results showed that the protein expression was significantly decreased after silencing SAPCD2 gene(P < 0.001).6.The results of CCK-8 assay showed that after silencing the expression of SAPCD2 in PC-3 and DU145 cells,the OD value decreased(P < 0.001)compared with the control group.7.The results of Transwell migration and Transwell invasion experiments showed that after silencing the expression of SAPCD2 in PC-3 and DU145 cells,the number of cell migration decreased compared with the control group(P < 0.001),and the number of invasion also decreased(P < 0.001).8.Flow cytometry results showed that after silencing the expression of SAPCD2 in PC-3 and DU145 cells,the proportion of apoptosis increased compared with the control group(P < 0.001).Conclusions:1.The expression of SAPCD2 gene in prostate cancer tissues was higher than that in normal prostate tissues.This study found that the high expression of SAPCD2 was correlated with Gleason score of more than 9(P < 0.05),and there was no correlation with other pathological data.2.Silencing SAPCD2 gene can significantly inhibit the proliferation,migration,invasion and apoptosis of prostate cancer cells(P<0.001). |
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