BACKGROUND: Ulcerative colitis (UC) is a significant type of inflammatory bowel disease (IBD). Ferroptosis is a type of procedural death that is associated with different types of diseases to various degrees. This study aimed to explore the key ferroptosis-associated genes in UC and assess their potential implications for biological therapy. METHODS: Bioinformatic prediction combined with experimental verification were applied to this study for the correlation between ferroptosis and UC. The Gene Expression Omnibus database was used to obtain disease-related gene expression data and then the identified genes expression were validated using Dextran Sulfate Sodium Salt (DSS) induced mouse colitis model. Next, the infiltration of immune cells was compared between UC and normal groups, and the correlation between different immune cells and the identified genes was analyzed. RESULTS: In this study, 91 ferroptosis-associated genes associated with UC were identified. Four genes (TIMP1, LPIN1, SOCS1, and CD44) were captured with diagnostic values among them, whose expression in DSS-induced mouse colitis was higher than control tissues. Besides, the expression levels of these four genes in the biological agents responsive group showed a distinction from those in active UC patients. Immune infiltration analysis revealed that there were positive or negative correlations between four genes with neutrophils, M0 and M1 macrophages, B cells, and NK cells. CONCLUSIONS: This study clearly indicates that immune cell infiltration in UC patients is associated with ferroptosis. The key genes associated with ferroptosis may have diagnostic value and predict therapeutic effects, which also can be helpful for discovering the molecular mechanism of UC.