Objective <p>Identifying novel molecular drivers is crucial to improving therapeutic strategies for hepatocellular carcinoma (HCC). This study aimed to investigate the clinical significance and biological function of UDP-glucuronate decarboxylase 1 (UXS1) in HCC.</p> Methods <p>UXS1 expression and its prognostic value were analyzed using the Gene Expression Profiling Interactive Analysis (GEPIA) database. In vitro, UXS1 was silenced in Huh-7 and HCCLM3 cell lines via siRNA. Malignant phenotypes were evaluated using CCK-8, Transwell, and wound healing assays. Transcriptome sequencing (RNA-seq) was performed to identify downstream mechanisms. Additionally, oxidative stress markers (catalase, superoxide dismutase, and glutathione) and key ferroptosis regulators (GPX4, SLC7A11) were assessed. Notably, rescue experiments using the ferroptosis inhibitor Ferrostatin-1 were conducted to validate the mechanism.</p> Results <p>UXS1 mRNA expression was significantly upregulated in HCC tissues, and high expression correlated with poor overall and progression-free survival. Functionally, UXS1 knockdown significantly inhibited HCC cell proliferation, migration, and invasion. Transcriptome analysis of 425 differentially expressed genes (DEGs) highlighted the enrichment of reactive oxygen species (ROS) and ferroptosis-related pathways. Mechanistically, UXS1 silencing disrupted ROS homeostasis and downregulated the anti-ferroptotic proteins GPX4 and SLC7A11. Significantly, treatment with Ferrostatin-1 effectively reversed the tumor-suppressive effects of UXS1 knockdown.</p> Conclusion <p>High expression of UXS1 serves as a prognostic biomarker for HCC. UXS1 knockdown suppresses HCC malignant progression, at least partially, by inducing ferroptosis via the SLC7A11/GPX4 axis. These findings suggest UXS1 is a potential therapeutic target and a novel ferroptosis checkpoint in HCC.</p>