Aucubin directly targets β-catenin to co-suppress Wnt and HIF-1 pathways in ovarian cancer: Computational and experimental validation
OV is a highly lethal malignancy plagued by chemoresistance, necessitating the development of novel therapeutic agents. The Wnt/β-catenin pathway, driven by its central component β-catenin (CTNNB1), is a key oncogenic axis in OV, making it an attractive therapeutic target. We employed an integrated strategy combining in vitro assays (MTT, Western blot), in silico analyses (network pharmacology, WGCNA, molecular docking, molecular dynamics simulations), and biophysical validation assays (CETSA, DARTS) to systematically investigate the anti-cancer mechanism of the natural compound AU. AU potently inhibited the viability of A2780 OV cells by inducing apoptosis. Our comprehensive bioinformatic analysis identified CTNNB1 as a high-confidence direct target of AU. Clinical data confirmed that CTNNB1 is overexpressed in OV and correlates with poor patient prognosis. Molecular docking and dynamics simulations predicted a stable AU-CTNNB1 interaction, which was then experimentally validated by both CETSA and DARTS, confirming direct target engagement in a cellular context. Mechanistically, AU treatment resulted in the dose-dependent suppression of key proteins in the Wnt/β-catenin and HIF-1 signaling pathways. AU exerts its anti-OV activity by directly binding to CTNNB1. This interaction inhibits the oncogenic Wnt/β-catenin pathway, leading to the concurrent suppression of the HIF-1 pathway and the induction of apoptosis. Our study provides a complete mechanistic rationale for the development of AU as a novel targeted therapy for ovarian cancer.