The aim of this study was to quantify the expression levels of the immune checkpoint molecules TIM-3 and Gal-9 in CRC tissues and adjacent non-tumor tissue, and to investigate their associations with key oncogenic mutations, including KRAS, NRAS, BRAF, PIK3CA, and AKT1, as well as MSI status. Here, LGALS9 is linked to colorectal carcinoma.