Abstract：Objective: To ide.pngy the expression of ubiquitin specific peptidase 3 (USP3) in bladder cancer and explore the possibility of USP3 serving as a prognostic marker for bladder cancer. Methods: Oncomine database was used to compare the expression of USP3 between bladder cancer and normal tissues. Furthermore, we further confirmed the USP3 expression in bladder cancer through detecting the USP3 level in our bladder cancer tissues and adjacent tissues of cancer as well as in bladder cancer cell lines and normal urothelial cell line using immunohistochemistry, qRT-PCR and Western blotting. Then we used TCGA database to analyze the relationship between the expression of USP3 and the prognosis of bladder cancer patients. Finally, USP3 association networks were predicted by STRING database and The analysis of gene ontology enrichment and KEGG Pathway was performed on the USP3 interaction proteins were. Results: All the analyses in Oncomine demonstrated that the level of USP3 was higher in the bladder cancer tissues than in normal tissues (P<0.05). The results of immunohistochemistry and qRT-PCR te.pngied that the protein and mRNA levels of USP3 were significantly overexpressed in bladder cancer tissues as compared with those in adjacent tissues of cancer (P<0.05). At the cellular level, the results of Western blotting and qRT-PCR also illustrated that the protein and mRNA levels of USP3 in two bladder cancer cell lines were significantly higher than in the normal urothelial cell line SV-HUC-1 (P<0.05). The survival analyses through the TCGA database revealed that high transcription levels of USP3 were associated with poorer overall survival in bladder cancer patients (P<0.05), but not with recurrence (P>0.05). Ten USP3-interacting proteins were predicted by the STRING database and the GO analysis revealed that these proteins were enriched in many biological processes, such as histone deubiquitination, chromatin organization and histone modification. They also participated in systemic lupus erythematosus, alcoholism, viral carcinogenesis and basal transcription factors. Conclusions: USP3 is up-regulated in bladder cancer and is closely associated with prognosis, suggesting USP3 may participate in bladder cancer progression. USP3 is valuable to be a prognostic marker of bladder cancer.
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