Background. The Distal-Less homeobox (DLX) gene family plays an important role in several tumors. However, the expression pattern, prognostic and diagnostic value, possible regulatory mechanisms, and the relationship between DLX family and immune in ltration in colon cancer (COAD) have not been systematically reported.Methods. We used Wilcoxon rank sum test and t-test to assess DLX gene family expression between COAD tissues and unpaired normal colon tissues, cBioPortal to analyze DLX gene family variants, R (version 3.6.3) to analyze DLX gene expression in COAD and the relationship between DLX gene family expression and clinical features and correlation heat map, the survival package [version 3.2-10] and the Cox regression module to assess the prognostic value of the DLX gene family, the pROC package [version 1.17.0.1] to analyze the diagnostic value of the DLX gene family, the R ( version 3.6.3) to analyze the possible regulatory mechanisms of DLX gene family members and related genes, the GSVA package [version 1.34.0] to analyze the relationship between the DLX gene family and immune in ltration, and the ggplot2 [version 3.3.3], and survminer package [version 0.4.9] and clusterPro ler package [version 3.14.3] for visualization. Results. DLX 2/3/4/5/6 were signi cantly upregulated in COAD patients. The expression of DLX family was associated with M stage, pathologic stage, primary therapy outcome, residual tumor, lymphatic invasion, T stage, N stage, age, perineural invasion, and history of colon polyps. DLX2/5 were independently correlated with the prognosis of COAD in multivariate analysis. DLX1/2/3/4/5/6 were involved in the development and progression of COAD by participating in immune in ltration and pathways, including breast cancer, gastric cancer, Hippo signaling pathway, Wnt signaling pathway, signaling pathways regulating pluripotency of stem cells, basal cell carcinoma, melanoma, and staphylococcus aureus infection. Conclusion. The DLX gene family can be used as potential diagnostic or prognostic biomarkers and therapeutic targets for COAD.