10-Hydroxy-2-decenoic acid, a bioactive component of royal jelly, induces p53-Mediated apoptosis in cholangiocarcinoma in vitro and in vivo.

The poor prognosis and limited treatment options for cholangiocarcinoma (CCA) continue to drive the search for novel therapeutics with clear mechanisms of action. 10-hydroxy-2-decenoic acid (10-HDA) is a unique bioactive fatty acid isolated from royal jelly, yet its therapeutic potential in CCA is unknown. In this study, we evaluated the anti-CCA properties of 10-HDA across cell-based, computational, transcriptomic, and animal models. Cell viability and clonogenic assays showed that 10-HDA strongly suppressed HUCCT1 and RBE cell growth by triggering extensive apoptosis. We also observed a marked decline in cell migration and invasion; rather than a purely targeted anti-metastatic effect, transcriptomic data suggested this motility loss is tightly linked to the drug's primary cytotoxicity and a parallel disruption of actin cytoskeleton dynamics. Computational predictions initially pointed toward apoptosis networks as primary targets, which was corroborated by our RNA-seq data highlighting a strong enrichment in both general apoptotic and specific TNF signaling pathways. Protein-level validation confirmed a dual-activation mechanism: 10-HDA upregulates the intrinsic p53/Bax axis while simultaneously engaging the extrinsic TNF/Caspase-8 cascade, both converging on Caspase-3 cleavage. These mechanisms translated effectively in vivo, where 10-HDA administration severely restricted xenograft tumor growth in nude mice without noticeable systemic toxicity. These findings identify 10-HDA as a potent, defined natural agent that halts CCA progression by activating comprehensive apoptotic programs.