NR4A1 Exerts Pro‐Tumor Role in Glioblastoma via Inducing xCT/GPX4‐Regulated Ferroptosis

Purpose This study investigates NR4A1's paradoxical roles in glioblastoma (GBM) progression, focusing on its mechanistic link to ferroptosis regulation. We aimed to resolve conflicting reports of NR4A1 as both an oncogene and a tumor suppressor by defining its transcriptional control over xCT/GPX4‐mediated iron homeostasis and its clinical relevance in glioma survival. Methods TCGA cohort analysis ( n = 163) correlated NR4A1 expression with survival endpoints (OS/PFI/DSS, log‐rank p < 0.05). Functional validation employed U87/U251 GBM models for viability (CCK‐8), proliferation (EdU/colony formation), and migration assays (Transwell/wound healing). RNA sequencing (DESeq2, FDR < 0.05) and ChIP‐qPCR identified NR4A1‐xCT transcriptional regulation. Ferroptosis was quantified via lipid peroxidation (MDA/GSH/Fe 2+ ELISA, C11 BODIPY), while Western blotting mapped the NR4A1/xCT/GPX4/P53 axis. Orthotopic xenografts ( n = 6/group) evaluated therapeutic efficacy using biweekly tumor volumetry. All data were analyzed in triplicate (GraphPad Prism 8.0; t ‐test/ANOVA, * p < 0.05). Conclusion NR4A1 drives GBM progression by transcriptionally activating xCT/GPX4 to suppress ferroptosis. Dual targeting of NR4A1 and ferroptosis pathways synergistically inhibits tumor growth (64% reduction vs. controls, p = 0.008), providing a mechanistic rationale for overcoming therapy resistance in GBM.