Homoharringtonine suppresses gastric cancer progression by targeting PI3K/ AKT/FOXO pathway and impairing the GSH-GPX4 antioxidant axis.

PURPOSE: To investigate the unexplored therapeutic potential and identify the specific molecular targets of homoharringtonine (HHT) in gastric cancer (GC). METHODS: HHT’s efficacy was evaluated in GC cell lines (MKN1/MKN45), normal epithelial cells (GES-1), and an HGC27 xenograft model. Mechanisms were investigated using RNA-sequencing, microscale thermophoresis (MST), molecular docking, and biochemical assays. RESULTS: HHT suppressed GC cell proliferation and migration, inducing G1-phase arrest and apoptosis, while demonstrating lower cytotoxicity toward normal GES-1 cells. MST and docking assays confirmed that HHT directly and stably binds to AKT (KD= 6.496 nM) and suppresses the PI3K/AKT/FOXO/GSK3β signaling pathway. Concurrently, transcriptomic profiling and experimental validation showed that HHT disrupts glutathione metabolism. It depleted intracellular GSH and downregulated GPX4, triggering ferroptosis-like oxidative damage and mitochondrial cristae loss. In vivo, HHT inhibited GC xenograft tumor growth without causing systemic toxicity, which was accompanied by decreased Ki-67 and increased Caspase-3 expression. CONCLUSION: HHT inhibits GC progression by suppressing AKT-mediated tumor growth and inducing GPX4-dependent ferroptosis. These findings highlight HHT as a promising targeted therapeutic for GC.