KIF24 depletion induces Fe–S-dependent mitochondrial dysfunction and AMPK/p53/SLC7A11 mediated ferroptosis in gastric cancer

Abstract Gastric cancer remains a major cause of cancer-related mortality, and ferroptosis has emerged as a therapeutically exploitable vulnerability in malignant cells. However, the upstream regulators that sustain ferroptosis resistance in gastric cancer remain incompletely defined. Here, we identify KIF24 as a previously unrecognized regulator of ferroptosis resistance in gastric cancer. Integrative transcriptomic analysis and single-cell profiling nominated KIF24 as a candidate ferroptosis-associated oncogenic factor enriched in malignant epithelial cells, and clinical validation confirmed its upregulation in gastric cancer tissues. Functional assays showed that KIF24 depletion suppressed gastric cancer cell proliferation, colony formation and xenograft tumor growth, while markedly increasing ferroptosis-associated phenotypes, including intracellular Fe²⁺ accumulation, GSH depletion, lipid peroxidation, mitochondrial ROS production and ferroptotic mitochondrial morphology. Mechanistically, KIF24 loss impaired Fe–S-dependent mitochondrial function, as reflected by reduced mitochondrial aconitase activity and altered mitochondrial Fe–S-related homeostasis. This mitochondrial stress was accompanied by AMPK activation, p53 phosphorylation and repression of SLC7A11, leading to disruption of the GSH-dependent antioxidant defense system. Pharmacological inhibition of ferroptosis, iron chelation, AMPK inhibition or p53 silencing partially rescued KIF24 depletion-induced ferroptotic injury, supporting the functional involvement of the AMPK–p53–SLC7A11 axis. In addition, preliminary structure-based virtual screening identified candidate compounds with potential KIF24-targeting activity, providing a basis for future therapeutic development. Together, our findings reveal a KIF24–mitochondrial stress AMPK–p53–SLC7A11 signaling axis that protects gastric cancer cells from ferroptosis and highlight KIF24 as a potential target for ferroptosis-based gastric cancer therapy.