Background and objectives: Colorectal cancer (CRC) is a disease that begins exclusively in the colon or rectum and is caused by the abnormal growth of glandular epithelial cells within the colon. Homocysteine, vitamin B6, and vitamin D are recognized to play a significant role in colorectal tumor pathogenesis. The aim of this study to investigate serum levels of Hcy, vitamin D and vitamin B6 in colorectal cancer. Methods: The current study investigated the vital roles of homocysteine, vitamin B6 and vitamin D in cancer development by comparing serum levels of colorectal cancer patients with those of the healthy control group at Nanakaly hospital. The levels of serum homocysteine (Hcy), vitamin B6 and vitamin D in 50 colorectal cancer patients and 50 from the healthy control group have been examined. Results: The result shows significantly high values of serum homocysteine (p<0.0001) and significant decreases in values of vitamin B6 and vitamin D (p<0.0001and 0.0010) respectively, in patients with CRC when compared with the healthy group. Conclusions: Our findings demonstrate that increased levels of homocysteine and decreased levels of vitamin B6 and vitamin D could be associated with the pathogenesis and progression of colorectal cancer.