Xin yixue (Jul 2023)
Expression level of RNF187 and its effect on the proliferation and invasion of prostate cancer cells
Abstract
Objective To investigate the expression level of RING finger protein 187 （RNF187） and its effect on the proliferation and invasion of prostate cancer （PCa） cells. Methods The differences in the expression of RNF187 between PCa and normal tissues was mined through TCGA and GEO databases. Normal prostate epithelial cell line RWPE-1， and human PCa cell lines LNCaP， C4-2， DU145 and PC-3 cell lines were cultured in vitro. The expression levels of RNF187 mRNA in RWPE-1， LNCaP， C4-2， DU145 and PC-3 were determined by quantitative fluorescent real-time PCR （RT-qPCR）. The expression level of RNF187 protein was determined by Western blot. DU145， PC-3 cells were transfected with RNF187-siRNA1 and RNF187-siRNA2. The relative expression level of RNF187 mRNA was determined by RT-qPCR. Cell viability was detected by CCK-8 assay. Cell proliferation was measured by 5-ethynyl-2´-deoxyuridine （EdU） labeling. Cell migration and invasion were measured by Transwell chamber. The expression levels of RNF187， epithelial-mesenchymal transition （EMT） marker proteins and matrix metalloproteinase-9 （MMP-9） were determined by Western blot. Results In TCGA and GEO datasets， the expression level of RNF187 in the PCa tissues was significantly up-regulated than that in the normal prostate tissues. Patients with higher Gleason scores and higher staging had higher expression levels of RNF187 （all P < 0.05）. The expression levels of RNF187 mRNA and protein in PC-3， LNCaP， C4-2 and DU145 cells were significantly up-regulated than those in RWPE-1 cells （all P < 0.01）. The relative expression levels of RNF187 mRNA in DU145 and PC-3 cells in the RNF187-si1 and RNF187-si2 groups were significantly down-regulated than those in the control group （all P < 0.001）. After RNF187 knock-down， the viability， proliferation， invasion and migration capabilities of DU145 and PC-3 cells were significantly decreased， the relative expression levels of RNF187， N-cadherin， Vimentin and MMP-9 proteins were significantly down-regulated， whereas the relative expression levels of E-cadherin were significantly up-regulated （all P < 0.05）. Conclusion RNF187 is over-expressed in PCa tissues. Knock-down of RNF187 can reduce the cell viability， proliferation， invasion and migration of DU145 and PC-3 cells.
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