AIF promotes ovarian cancer progression by competitively binding TRAP1 to stabilize MtCK1 to drive phosphocreatine synthesis

Ovarian cancer is a common malignant tumor in women, characterized by delayed diagnosis, high metastatic potential, frequent drug resistance, and poor clinical prognosis. Although aberrant expression of apoptosis-inducing factor (AIF) has been reported in several human cancers, its role in ovarian cancer has not yet been fully elucidated. Therefore, this study was performed to investigate the role of AIF in ovarian cancer progression. Integrated analyses of bulk RNA-sequencing, single-cell sequencing, and spatial transcriptomic data from public databases including TCGA and GEO, were performed, and significant overexpression of AIF in ovarian cancer tissues, together with a positive correlation with tumor grade, was confirmed. Cellular functional assays, including MTT, colony formation, Transwell, and wound healing, combined with subcutaneous tumorigenesis and intraperitoneal metastasis models in nude mice, were used to demonstrate the promoting effects of AIF on tumor proliferation, migration, and invasion. Furthermore, the effects of AIF on cellular metabolism, including oxidative phosphorylation, glycolysis, and ATP production were evaluated using metabolomic and metabolic assays. Mechanistic studies employing co-immunoprecipitation (Co-IP) and a proteasome degradation assay were performed to assess the interactions among AIF, TRAP1, and MtCK1, as well as their contribution to phosphocreatine synthesis. AIF was significantly overexpressed in ovarian cancer tissues and positively associated with tumor grade. Functional studies confirmed that AIF promoted tumor proliferation and metastasis both in vitro and in vivo. Moreover, oxidative phosphorylation and glycolysis in ovarian cancer cells were markedly enhanced, accompanied by increased ATP production. Mechanistically, AIF drove metabolic reprogramming through competitively binding to TRAP1, thereby promoting MtCK1 stabilization and enhancing phosphocreatine synthesis. Further analyses demonstrated that the AIF-TRAP1-MtCK1 axis was essential in the maintenance of oncogenic phenotypes, and the effects induced by AIF knockdown were effectively reversed by phosphocreatine supplementation. Collectively, these findings identified AIF-mediated phosphocreatine metabolic reprogramming as a potential therapeutic target in ovarian cancer.