biostudies-literatureUnknown14(2)Wang NThe significance of 5-methylcytosine (m⁵C) methylation in human malignancies has become an increasing focus of investigation. Here, we show that m⁵C regulators including writers, readers and erasers, are predominantly upregulated in urothelial carcinoma of the bladder (UCB) derived from Sun Yat-sen University Cancer Center and The Cancer Genome Atlas cohort. In addition, NOP2/Sun RNA methyltransferase family member 2 (NSUN2) as a methyltransferase and Aly/REF export factor (ALYREF) as a nuclear m⁵C reader, are frequently coexpressed in UCB. By applying patient-derived organoids model and orthotopic xenograft mice model, we demonstrate that ALYREF enhances proliferation and invasion of UCB cells in an m⁵C-dependent manner. Integration of tanscriptCell death & disease139https://www.ebi.ac.uk/biostudies/studies/S-EPMC9938871biostudies-literaturem⁵C-dependent cross-regulation between nuclear reader ALYREF and writer NSUN2 promotes urothelial bladder cancer malignancy through facilitating RABL6/TK1 mRNAs splicing and stabilization.PMC9938871Zhou ZHChen RXLiu ZWWen JHNing KZhou FJLi YHLi XDWang NYe YLWei WSZhang XPDeng MHDong Bfalsem⁵C-dependent cross-regulation between nuclear reader ALYREF and writer NSUN2 promotes urothelial bladder cancer malignancy through facilitating RABL6/TK1 mRNAs splicing and stabilization.The significance of 5-methylcytosine (m⁵C) methylation in human malignancies has become an increasing focus of investigation. Here, we show that m⁵C regulators including writers, readers and erasers, are predominantly upregulated in urothelial carcinoma of the bladder (UCB) derived from Sun Yat-sen University Cancer Center and The Cancer Genome Atlas cohort. In addition, NOP2/Sun RNA methyltransferase family member 2 (NSUN2) as a methyltransferase and Aly/REF export factor (ALYREF) as a nuclear m⁵C reader, are frequently coexpressed in UCB. By applying patient-derived organoids model and orthotopic xenograft mice model, we demonstrate that ALYREF enhances proliferation and invasion of UCB cells in an m⁵C-dependent manner. Integration of tanscript2023-01-01T00:00:00Z2023 Feb2025-04-04T20:10:04.838Z2025-02-18T22:50:53.027ZS-EPMC99388713680625310.1038/s41419-023-05661-y