Abstract
Laryngeal cancer (LC) is one of the malignant tumors affecting the head and neck region. 5-methylcytosine (m5C) is a common RNA modification that contributes to the carcinogenesis process. Aly/REF export factor (ALYREF), an m5C reader protein, exhibits dysregulation in various cancers, yet its role in LC remains unclear. To investigate this, we utilized CCK8 assays, colony formation, and flow cytometry for phenotypic analysis, while western blotting and real-time quantitative PCR were employed to assess ALYREF expression. The molecular mechanisms were explored using methylated RNA immunoprecipitation (MeRIP), RIP, dual-luciferase reporter assays, and protein stability experiments. Our data indicated that ALYREF was substantially overexpressed in LC tissues and cells. Silencing ALYREF resulted in reduced cell viability and proliferation in vitro, enhanced apoptosis, and suppressed tumor growth in vivo. Mechanistically, ALYREF interacted with DDX11 mRNA and enhanced its stability through NSUN2-mediated m5C modifications. Overall, our study suggested that ALYREF drove LC progression by stabilizing DDX11 mRNA. The findings provided novel insights into the role of ALYREF-induced m5C modifications in LC, opening up new perspectives for further research.