【Abstract】 Objective To investigate expression and clinical significance of the carcinoembryonic antigen (CEA) and glutaminase type 2 (GLS2) in rectal cancer. Methods 115 patients with rectal cancer were enrolled in the study. All patients underwent laparoscopy. The specimens of rectal cancer were selected. The expression levels of CEA and GLS2 in the specimens were determined by immunohistochemistry. The correlation of their expression and clinical significance in rectal cancer was further explored. Results There were 60 patients (521%) with high expression of CEA and 74 cases (643%) with high expression of GLS2 in rectal cancer tissues. There was a significant correlation between CEA and GLS2 positive expression. The CEA expression was related with patient age, tumor stage and lymph node metastasis in rectal cancer tissues. The expression of GLS2 is related to patient age, lymph node metastasis and tumor long diameter in rectal cancer tissues. The prognosis of patients with rectal cancer was affect by the expression of CEA and GLS2, age, tumor diameter, tumor stage and lymph node metastasis. Conclusion CEA and GLS2 are higher expressed in rectal cancer tissues and are related to the age and lymph node metastasis of the patients. The combined detection of these two factors is helpful for the diagnosis and treatment of rectal cancer.