NSUN2-mediated R-loop stabilization as a key driver of bladder cancer progression and cisplatin sensitivity

R-loops are critical structures that play pivotal roles in regulating genomic stability and modulating gene expression. This study investigates the interactions between the 5-methylcytosine (m 5 C) methyltransferase NOP2/Sun RNA methyltransferase 2 (NSUN2) and R-loops in the transcriptional dynamics and damage repair process of bladder cancer (BCa) cells. We observed markedly elevated levels of R-loops in BCa cells relative to normal urothelial cells. NSUN2 was identified as a regulator of R-loops, acting to bind and stabilize their structure through a process dependent on its m 5 C catalytic activity. The histone methyltransferase Enhancer of Zeste Homolog 2 (EZH2) was found to interact with NSUN2. Our results demonstrated that NSUN2 facilitates the epigenetic silencing of the tumor suppressor gene PR Domain Zinc Finger Protein 11 (PRDM11) by recruiting EZH2, thereby advancing the progression of BCa. Furthermore, NSUN2 knockdown sensitizes tumors to cisplatin, resulting in reduced tumor growth and increased DNA damage levels, which was associated with reduced recruitment of MRE11 to damage sites, thereby impairing homologous recombination repair. These findings enhance our understanding of BCa pathogenesis and identify new potential targets for therapeutic intervention. • R-loops are elevated in bladder cancer cells compared to normal urothelial cells. • NSUN2 regulates R-loops via m 5 C catalytic activity, stabilizing their structure. • NSUN2 promotes cancer growth by silencing PRDM11 through interaction with EZH2. • NSUN2 knockdown sensitizes bladder cancer cells to cisplatin by reducing R-loops.