<p>Chemoresistance and ferroptosis inhibition in ovarian cancer (OC) may contribute to tumor progression. This study aimed to investigate the role of RUNX2 in regulating ferroptosis and cisplatin (CDDP) resistance in OC, along with its post-translational modification mechanisms. Parental and CDDP-resistant OC cell lines, as well as xenograft models in nude mice, were employed to explore the molecular functions of RUNX2. Cellular function and drug resistance were assessed using cell counting kit-8, colony formation, and flow cytometry. Ferroptosis activity was evaluated by measuring protein markers (FTH1, GPX4, and ASCL4), reactive oxygen species, Fe²⁺, GSH/GSSG, malondialdehyde, and lipid peroxidation levels. The palmitoylation of RUNX2 were assessed using acyl-biotin exchange, Click-iT pull-down, and palmitoyl acyltransferase assays. Based on the predicted sites, Flag-RUNX2 wild-type and mutant vectors were constructed, and Myc-ZDHHC14 was exogenously introduced to examine ZDHHC14-mediated RUNX2 palmitoylation. The results indicated that RUNX2 significantly suppressed ferroptosis and enhanced CDDP resistance in OC both in vitro and in vivo. Downstream, RUNX2 transcriptionally regulated YAP1 and activated the YAP1/GLS1 axis. Additionally, palmitoylation of RUNX2 was observed in OC cells. Bioinformatics analysis predicted the binding of RUNX2 with palmitoyltransferase ZDHHC14. Exogenous induction of Myc-ZDHHC14 drove RUNX2 palmitoylation, an effect that was abolished by the RUNX2-C13S mutation. Further in vitro and in vivo experiments confirmed that ZDHHC14-driven palmitoylation of RUNX2-C13S inhibited ferroptosis and chemosensitivity in OC by activating the YAP1/GLS1 axis. To conclude, ZDHHC14-driven S-palmitoylation of RUNX2 inhibits ferroptosis and CDDP sensitivity in OC by positively regulating the YAP1/GLS1 axis.</p>