Objective: To determine the correlation of MLH1 and MSH2 expressions with clinicopathologic characteristics in colorectal cancer (CC). Methods: Clinical data, CC tissue, and paracancerous tissue from 88 patients treated in Baoji City People’s Hospital from February 2015 to February 2017 were analyzed retrospectively. The relative expression levels of MLH1 and MSH2 in the tissues were measured with qRT-PCR, and the relationship of MLH1 and MSH2 with the pathological data of patients was analyzed. The value of MLH1 and MSH2 in the diagnosis of clinical stage, lymph node metastasis, and degree of differentiation in CC patients was analyzed by receiver operating curve (ROC). Cox regression analysis was applied to identify factors affecting prognosis. Results: The relative expression levels of MLH1 and MSH2 in CC tissue were lower than those in paracancerous tissue (P < 0.001). Tumor node metastasis stage (III + IV), poor differentiation, and lymph node metastasis were significantly increased in patients with low MLH1 and MSH2 expressions (P < 0.05). The levels of MLH1 and MSH2 in CC tissue of patients at stage I with moderately- or well-differentiated non-metastatic disease were higher than those in patients at stage II-IV with poor differentiation and lymph node metastasis, showing a good predictive ability. The 5-year survival rate of patients with low MLH1 and MSH2 expressions was lower as compared to its counterpart (P < 0.01). Conclusion: The low expressions of MSH2 and MLH1 in CC tissue have a correlation with pathological characteristics and survival, so they can be used as auxiliary references for the prognosis in CC patients.