Study On The Role And Mechanism Of High Expression Of RNF187 In Promoting Migration And Invasion Of Bladder Cancer

Background:Ring Finger Protein 187(RNF187;RACO-1)is a member of the E3 ubiquitinated ligases.Its overexpression has been shown to promote the malignant progression of a variety of tumours,but its role in bladder cancer(BC)and its mechanism are unclear.In this study,we investigated the effect of RNF187 on the biological function of bladder cancer cells and the effect of the level of RNF187 expression on the prognosis of clinical bladder cancer patients by using RNA sequencing to find out the molecular mechanism of action of RNF187 and validate it,which was combined with bioinformatics analysis to explore the mechanism of action of RNF187 in bladder cancer.Methods:Based on data from The Cancer Genome Atlas(TCGA)database and immunohistochemistry(IHC)results,the expression of RNF187 gene in tumor tissues of bladder cancer patients was identified.The lentiviral vectors RNF187-sh RNAZS-Green and RNF187-PCDH-GFP+Puro were constructed and infected with T24 and J82 cells and T24 and EJ cells,respectively,to validate the effects of RNF187 on tumour cell proliferation,by CCK-8,scratch assay,cell flowmetry and Transwell invasion assay at the in vitro level.The effect of RNF187 on tumour cell proliferation,apoptosis,migration and invasion was verified in vitro by CCK-8,scratch assay,cell flow assay and Transwell invasion assay.In vivo studies were performed in nude mice to verify the effect of RNF187 knockdown on tumour proliferation in vivo.The expression of RNF187 in different cell lines and bladder cancer tissues was verified by real-time fluorescence quantitative real-time PCR(RT-q PCR)and protein immunoblotting(Western blotting,WB),and the related signalling pathways and mechanisms were verified.Result:We found that the expression of RNF187 gene was significantly higher in tumour tissues of bladder cancer patients than in paraneoplastic tissues,and that high expression of RNF187 gene was associated with a poorer prognosis.When RNF187 expression was down-regulated,bladder cancer cells showed reduced proliferation,migration and invasion,and increased apoptosis,whereas the opposite was true for overexpression of RNF187.In vivo studies showed that knockdown of RNF187 delayed tumour growth and reduced tumour weight and size in nude mice,and WB analysis showed that RNF187 controlled bladder cancer cell migration and invasion through the PI3K/AKT signalling pathway by regulating the expression of ITG-β8(integrins)and ITG-β3.Conclusion:According to our results,RNF187 holds promise as a new molecular marker for predicting the prognosis of bladder cancer patients and as a potential therapeutic target.