<p>Ferroptosis has emerged as a potential therapeutic target for non-small cell lung cancer (NSCLC), but its regulatory mechanisms remain elusive. Protein tyrosine kinase 6 (PTK6) is overexpressed in NSCLC and linked to poor prognosis, though its role in ferroptosis is unknown. CCK-8 assay was performed to assess cell viability. Intracellular Fe²⁺ level was measured using an iron assay kit. Lipid peroxidation was evaluated using the C11 BODIPY probe. Dual-luciferase reporter and ChIP assays were employed to investigate FOXO3’s interaction with the APOL3 promoter. PTK6-YTHDF2 interaction was examined using Co-IP assay, and YTHDF2-FOXO3 interaction was detected using RIP assay. PTK6 knockdown exacerbated Erastin-induced ferroptosis in NSCLC cells. Mechanistically, PTK6 enhanced YTHDF2-mediated FOXO3 mRNA degradation by phosphorylating YTHDF2. FOXO3 silencing reversed PTK6 depletion’s pro-ferroptotic effects. FOXO3 transcriptionally activated APOL3 expression. APOL3 knockdown negated PTK6 silencing-driven ferroptosis sensitization. PTK6 inhibited NSCLC cells ferroptosis by promoting m6A-YTHDF2-dependent FOXO3 mRNA degradation through phosphorylating YTHDF2, thereby suppressing FOXO3-mediated APOL3 transcriptional activation.</p>