Elbasvir Triggers Ferroptosis in Esophageal Squamous Cell Carcinoma Through NCOA4-Mediated Ferritinophagy

Objective: Esophageal squamous cell carcinoma (ESCC) remains treatment-resistant; we explored Elbasvir, an NS5A inhibitor, as a ferroptosis inducer. Methods: Cell viability was assessed by CCK-8 assays. Apoptosis and cell cycle were analyzed via flow cytometry, and key markers via Western blotting. In vivo efficacy was evaluated using BALB/c nude mouse xenografts. Proteomic analysis was conducted by mass spectrometry. Ferroptosis induction was verified via TEM, JC-1, FerroOrange, DCFH-DA, MDA assays, and Western blotting of NCOA4, Ferritin, and FTH1. Binding to NCOA4 was confirmed by surface plasmon resonance (SPR) and drug affinity responsive target stability (DARTS) assays. Results: Elbasvir (40 μM, 48 h) suppressed KYSE150/TE1 viability, induced apoptosis/G0/G1 arrest, and inhibited xenograft growth without toxicity. Proteomics identified ferroptosis as the top pathway. SPR/DARTS confirmed NCOA4 binding. NCOA4 knockdown reduced ferroptosis; overexpression enhanced it. Elbasvir triggered NCOA4-mediated ferritinophagy, FTH1 degradation, iron accumulation, and lipid peroxidation. Discussion: Elbasvir targets NCOA4-FTH1 to induce ferroptosis, offering a repurposing strategy for ESCC. Its safety profile supports clinical translation, with potential applications in iron metabolism-dependent cancers.