Supplementary file 1_ALYREF regulates glycolysis and promotes the progression of endometrial cancer through m5C modification of PKM2 mRNA.docx

Here, we investigated the biological function and molecular mechanism of the RNA-binding protein ALYREF in EC. Bioinformatics analysis of public datasets revealed that ALYREF is significantly upregulated in EC tissues, and high ALYREF expression is associated with poor overall survival in the Kaplan-Meier Plotter cohort.

Clinical validation using paired EC tissues confirmed ALYREF upregulation at protein level, and high ALYREF expression was associated with aggressive clinicopathological features. Functional assays revealed that ALYREF promotes EC cell proliferation, migration, and invasion in vitro and enhances tumor growth and metastasis in vivo. Mechanistically, ALYREF increases the stability of PKM2 mRNA, which we demonstrate to be m⁵C-modified in EC, leading to elevated PKM2 protein expression, enhanced glycolytic metabolism and tumor progression.