N6-methyladenosine reader YTHDF2 facilitates malignant progression of gallbladder cancer by suppressing NME1 expression.

Gallbladder carcinoma (GBC) is a highly aggressive malignancy characterized by an extremely poor prognosis and limited therapeutic options. N6-methyladenosine (m6A), the most prevalent post-transcriptional modification in eukaryotic mRNA, plays a pivotal role in regulating tumor progression. Screening of an siRNA library targeting YTH-domain family reader proteins identified YTH domain-containing family protein 2 (YTHDF2) as essential for the malignant phenotype of GBC cells. YTHDF2 was aberrantly upregulated in GBC tissues and served as an independent predictor of poor prognosis. Genetic silencing of YTHDF2 markedly suppressed both proliferation and metastatic potential of GBC cells in vitro and in vivo. Mechanistically, YTHDF2 transcription is upregulated through P300 histone acetyltransferase-mediated H3K27ac modification. YTHDF2 promotes mRNA degradation by binding to m6A-modified sites within the transcript encoding the metastasis suppressor non-metastatic protein 1 (NME1), thereby repressing NME1 expression and accelerating GBC malignancy. Co-treatment with the P300 inhibitor C646 and the m6A modification inhibitor STM2457 synergistically inhibited YTHDF2-mediated GBC malignancy. Collectively, these findings establish the P300-YTHDF2-m6A-NME1 axis as a critical driver of GBC progression and indicate that combined C646 and STM2457 treatment represents a novel and promising therapeutic strategy for GBC.