Comprehensive analysis and functional validation of key genes in bladder cancer progression

Abstract Bladder cancer exhibits considerable heterogeneity, posing challenges for prognosis and treatment. In this study, we utilized an integrated approach involving single‐cell RNA sequencing and the Cancer Genome Atlas dataset analysis, combined with in vitro functional assays, to identify key genes associated with bladder cancer progression. A novel three‐gene signature was identified and validated for its prognostic relevance in independent patient samples. Among these genes, FAM3B emerged as the most significant driver, consistently overexpressed in high‐risk patients and strongly associated with poor survival outcomes. Functional assays in bladder cancer cell lines demonstrated that FAM3B promotes cell proliferation, clonogenicity, migration, and invasion. Additionally, the three‐gene signature showed a significant correlation with advanced tumor grade and stage, highlighting its potential utility for clinical risk stratification and personalized treatment strategies. These findings not only enhance our understanding of bladder cancer biology but also underscore the therapeutic potential of targeting FAM3B and the three‐gene signature as prognostic biomarkers.