α-Hederin inhibits osteosarcoma progression by triggering ferroptosis and disrupting mitochondrial function via PI3K/AKT signalling

Osteosarcoma is the most common primary malignant bone tumour in adolescents, with a poor prognosis and high rates of recurrence and metastasis. Ferroptosis, a form of regulated cell death associated with iron overload and lipid peroxidation, and mitochondrial dysfunction have been recognised as potential therapeutic targets in cancer. α-Hederin, a natural triterpenoid saponin, has shown antitumor effects in several cancers, but its role in osteosarcoma remains unclear. This study aimed to investigate the pharmacological effects and mechanisms of α-hederin on osteosarcoma. We integrated network pharmacology and molecular docking to identify potential targets and pathways, and validated the findings using <i>in vitro</i> experiments in 143B osteosarcoma cells. Network pharmacology analysis revealed 27 overlapping targets between α-hederin and osteosarcoma, with significant enrichment in the PI3K/AKT and ferroptosis pathways. Molecular docking confirmed strong binding affinity between α-hederin and key targets. Functional assays demonstrated that α-hederin inhibited cell proliferation and migration, increased intracellular Fe²⁺ and lipid reactive oxygen species levels, disrupted mitochondrial membrane potential and ATP production, and downregulated mitochondrial biogenesis proteins. Transmission electron microscopy further revealed typical mitochondrial morphological changes associated with ferroptosis. Western blotting showed decreased expression of GPX4 and SLC7A11, increased ACSL4 levels, and suppressed activation of the PI3K/AKT pathway. Moreover, these effects were partially reversed by the ferroptosis inhibitor Ferrostatin-1. Collectively, these results indicate that α-hederin exerts antitumor effects against osteosarcoma by inducing ferroptosis and impairing mitochondrial function, partly through inhibition of the PI3K/AKT signalling pathway, providing a potential therapeutic strategy for osteosarcoma.