UBA1 drives cell cycle to promote malignancy and serves as a therapeutic target in Ewing Sarcoma

Background Ewing sarcoma (ES) is an aggressive malignancy with high metastatic potential and poor prognosis, necessitating novel biomarkers and therapeutic targets. This study investigates the role of the ubiquitin-activating enzyme E1 (UBA1) in ES pathogenesis and its clinical relevance. Methods Bioinformatic analysis of GEO datasets was performed. UBA1 expression was validated in clinical samples and ES cell lines (A673, RDES) via Western blot and qPCR. To investigate the functional role and therapeutic potential of UBA1 in ES, we performed siRNA-mediated knockdown followed by a series of in vitro functional assays, including assessments of proliferation, colony formation, migration, invasion, cell cycle distribution, and EdU incorporation. Mechanistic studies were subsequently conducted through the WB experiment. In vivo experiments were carried out using a xenograft mouse model to evaluate the effect of UBA1 depletion on tumor growth. Additionally, the UBA1-specific inhibitor TAK-243 was used to assess the therapeutic potential of targeting UBA1. Results UBA1 expression was significantly elevated in ES tissues, and high UBA1 levels correlated with poor prognosis. Knockdown of UBA1 suppressed malignant phenotypes, including proliferation, colony formation, migration, and invasion, while inducing cell cycle arrest and inhibiting DNA synthesis. Mechanistically, UBA1 depletion led to dysregulation of the G2/M checkpoint pathway, consistent with Gene Set Enrichment Analysis (GSEA) predictions. In vivo experiments confirmed the oncogenic role of UBA1, and treatment with the UBA1 inhibitor TAK-243 recapitulated the effects observed upon UBA1 knockdown. Conclusion Targeting UBA1 disrupts cell cycle progression and inhibits proliferation in ES, underscoring its therapeutic potential.