Abstract: Objective To evaluate the short-term efficacy and toxicity of raltitrexed or fluorouracil in combination with irinotecan as the second-line treatment for advanced colorectal cancer. Methods Totally 52 advanced colorectal cancer patients with failed first-line chemotherapy were divided into two groups, Group A: 25 patients treated with raltitrexed plus irinotecan and Group B: 27 patients treated with irinotecan plus fluorouracil and leucovorin. The efficacy, toxicity and survival time of the treatments were compared between two groups. Results The effective rates were 36.0% in Group A and 11.5% in Group B(P﹤0.05). The disease control rates were 76% in Group A and 57.7% in Group B(P>0.05). The median time to disease progression were 6 months in Group A and 4.5 months in Group B(P>0.05). Conclusion Raltitrexed combined with irinotecan is effective and safe as the second-line treatment for advanced colorectal cancer, and further clinical application is recommended.