Rna M6a Methylation Promotes Tumor Development and Wasf3 Translation in Esophageal Squamous Cell Carcinoma

Background N6-methyladenosine (m6A) RNA modification coordinates the development and progression of several types of cancer, including esophageal squamous cell carcinoma (ESCC). ESCC tissues exhibit abnormal m6A modification and regulator levels, but the specific effects of this dysregulation on ESCC and its prognosis remain unclear. Methods Multi-omics sequencing data analysis was used to identify key targets in ESCC. Reverse transcription quantitative PCR (RT-qPCR), western blotting, immunohistochemistry, and immunofluorescence analyses were used to detect expression levels. Tissue microarrays were used to investigate prognostic potential. RNA pull-down, RNA immunoprecipitation (RIP)-qPCR, methylated RIP-qPCR, and co-immunoprecipitation assays were employed to validate interactions between key proteins and RNA. Polysome profiling and tethering assays were performed to assess translational efficiency. Lipid nanoparticle (LNP)-delivered small interfering RNA was used to evaluate treatment potential in cell line- and patient-derived xenograft models. Results WASF3 levels were significantly elevated in ESCC tissues, and ESCC patients with high WASF3 expression had significantly worse prognosis. WASF3 suppressed the proliferation of ESCC cells and inhibited colony formation and cell cycle progression. Mechanistically, METTL3 interacted with WASF3 and mediated m6A modification of its mRNA. IGF2BP2 enhanced WASF3 translation by binding to the m6A site in its 3′ untranslated region, and highly expressed WASF3 activated the MAPK signaling pathway by interacting with phosphorylated p38 (p-p38), thereby promoting ESCC progression. Finally, LNP small interfering RNA targeting WASF3, both alone and in combination with paclitaxel, could successfully suppress ESCC tumorigenesis. Conclusion Our findings demonstrate that WASF3 plays a pivotal role in ESCC and highlight the functional significance of the METTL3/m6A/WASF3/IGF2BP2 axis in regulating ESCC progression, which could facilitate the development of novel prognostic and therapeutic targets for ESCC.