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Background: Many pseudogenes possess biological activities and play important roles in the pathogenesis of various types of cancer including bladder cancer (BlCa), which still lacks suitable molecular biomarkers. Recently, pseudogenes were found to be significantly enriched in a pan-cancer classification based on the Cancer Genome Atlas gene expression data. Among them, the top-ranking pseudogene was the proliferation-associated 2G4 pseudogene 4  (PA2G4P4 ). Methods: Genomic and transcript features of  PA2G4P4  were determined by GeneBank database analysis followed by 5’ RACE experiments. Therefore, we conducted a retrospective molecular study on a cohort of 45 patients of BlCa.  PA2G4P4  expression was measured by RT-qPCR, whereas  PA2G4P4  transcript distribution was analyzed by in situ hybridization on both normal and cancerous histological sections and compared to the immunolocalization of its parental  PA2G4/EBP1  protein. Finally, we tested the effects of  PA2G4P4  depletion on proliferation, migration, and death of BlCa cells. Results: We showed for the first time  PA2G4P4  overexpression in BlCa tissues and in cell lines.  PA2G4P4  distribution strictly overlaps PA2G4/EBP1 protein localization. Moreover, we showed that  PA2G4P4  knockdown affects both proliferation and migration of BlCa cells, highlighting its potential oncogenic role. Conclusions:  PA2G4P4  may play a functional role as an oncogene in BlCa development, suggesting it as a good candidate for future investigation and new clinical applications.

Role of PA2G4P4 pseudogene in bladder cancer tumorigenesis