The Role Of RCAS1/EBAG9 In The The Process Of Epithelial Mesenchymal Transition In Prostate Carcinoma

ObjectiveTo determine the expression of RCAS1/EBAG9 in prostate carcinoma tissues,and its relationship with the clinical relevance in prostate carcinoma.By detecting the different expression of RCAS1/EBAG9 in prostate carcinoma cells,RCAS1 high expression of recombinant plasmid and RCAS1/EBAG9 shRNA plasmid were transformed into prostate carcinoma cells separately to detect the expression of epithelial-mesenchymal transition markers and RCAS1/EBAG9,explaining the possible effect of RCAS1/EBAG9 on epithelial-mesenchymal transition of prostate carcinoma.Methods1.Immunohistochemical staining were conducted separately between 50 cases of postoperative prostate carcinoma(PCa)tissue samples and 18 cases of benign prostatic hyperplasia(BPH)tissue samples to detect the expression of RCAS1/EBAG9.The expression levels of RCAS1/EBAG9 were determined in 8 cases with prostate tissues using RT-qPCR and Western blot methods(4 cases of PCa and 4 cases of BPH),the results were analysed by statistical methods.The prostate carcinoma tissues were classified according to the pathological grades(well differentiation and poorly differentiation)and clinical stages(T1+T2 stages and T3+T4 stages),to analyse the relationship between the expression of RCAS1/EBAG9 and prostate carcinoma clinical relevance.2.The RCAS1/EBAG9 shRNA plasmid shRCAS1-406 and the RCAS1/EBAG9 high expression of recombinant plasmid were obtained.Our precendent experimental work have revealed the different expression levels of RCAS1/EBAG9 in prostate carcinoma cell lines C42,LNCaP,PC3 and DU145.The RCAS1/EBAG9 high expression plasmid,negative control(NC),blank control were transferred into prostate carcinoma cells PC3 and DU145 separately.Similarly,the shRCAS1-406 plasmid and its negative control group were transferred into C42 and LNCaP.RT-qPCR and Western blot were performed to identify the differences between the expression of RCAS1/EBAG9 and the epithelial-mesenchymal transition markers(such as E-cadherin,N-cadherin,Vimentin)to confirm the possible effect of RCAS1/EBAG9 on the epithelial-mesenchymal transition process.Results1.The immunohistochemical staining results suggested that the expression levels of RCAS1/EBAG9 in prostate carcinoma tissues and benign prostate tissues were 74%(37/50),11.1%(2/18)(P<0.05).RT-qPCR and Western blot results showed that the expression levels of RCAS1/EBAG9 in prostate carcinoma tissues were significantly higher than those in benign prostate tissues.The expression of RCAS1/EBAG9 had a positive relationship with clinical stages,and the expression of RCAS1/EBAG9 in the poorly differentiated groups were significantly higher than those in the well differentiated groups in prostate carcinoma.2.By transferring RCAS1-high plasmid and shRCAS1-406 plasmid into the different prostate carcinoma cells according to their relative expression levels of RCAS1/EBAG9,RT-qPCR and Western blot revealed that the expression levels of RCAS1/EBAG9 with the high expression plasmid in the prostate carcinoma cells PC3 and DU145 were significantly higher than those in the other two groups,and the expression of epithelial cell marker E-cadherin was consisted with the expression of RCAS1/EBAG9,while the expression levels of mesenchymal cell marker N-cadherin and Vimentin were negative correlation with the expression of RCAS1/EBAG9.Similarly,the expression of RCAS1/EBAG9 with the shRCAS1-406 plasmid groups in the prostate carcinoma cells C42 and LNCaP were obviously less than those in the negative control groups,and the epithelial cell marker E-cadherin had a positive correlation with the expression of RCAS1/EBAG9,while the mesenchymal cell markers N-cadherin and Vimentin had a negative correlation.Conclusions1.The expression levels of RCAS1/EBAG9 in prostate carcinoma tissues are significantly higher than that in benign prostatic hyperplasia tissues and are associated with prostate carcinoma clinical stages and pathological grades.2.RCAS1/EBAG9 is involved in the process of epithelial mesenchymal transition in prostate carcinoma and may be associated with the metastasis of prostate carcinoma.