Proteomic Analysis Reveals AMPKα ‐Mediated Antiandrogen Resistance via Ferroptosis Suppression in Prostate Cancer

Metastatic prostate cancer often develops resistance to second‐generation androgen receptor pathway inhibitors (ARPIs) such as enzalutamide and darolutamide, limiting their long‐term efficacy. In this study, we applied the high‐precision absolute quantitative proteomics platform iMPAQT to comprehensively profile protein expression changes in multiple prostate cancer cell lines and their ARPI‐resistant derivatives. Our analysis revealed consistent dysregulation of ferroptosis suppressor pathway alongside elevated phosphorylation of AMPKα signaling in resistant cells. While antiandrogen treatment increased lipid peroxidation in parental cells, resistant cells adapted by upregulating key ferroptosis‐regulating proteins including FTH1 and GPX4, thereby evading ferroptosis. Functional assays demonstrated that the GPX4 inhibitor RSL3 selectively suppressed proliferation of resistant cells more potently than parental cells. Moreover, combination treatment with AMPKα inhibitors enhanced the anti‐proliferative effect of RSL3. These findings suggest that ferroptosis suppression is a critical mechanism underpinning resistance to ARPIs in prostate cancer. Targeting pathways involved in ferroptosis regulation in combination with AMPKα inhibitors holds promise for overcoming therapeutic resistance and improving patient outcomes.