Objective: To evaluate the efficacy and toxicity of the combination chemotherapy of docetaxel, oxaliplatin and fluorouracil (DOF regimen) on advanced gastric cancer. Methods: Sixty-five patients with advanced gastric cancer were randomized into DOF regimen group(n = 33) and FOLFOX4 regimen group (n = 32). Clinical efficacy and adverse reactions were observed in the two groups. Results: The objective response rate was 51.5% in DOF regimen group. The median TTP in DOF regimen group was 8.6 months and the one year survival rate was 57.3%. In FOLFOX4 regimen group, the objective response rate was 46.9%%. The median TTP was 5.8 months and the one year survival rate was 41.2%. Statistical analysis indicated that the two groups had similar objective response rate (P > 0.05); TTP and one year survival rate in DOF regimen group were better than FOLFOX4 regimen group(P <0.05); The incidences of neutropenia, thrombocytopenia and hair loss were higher in DOF regimen group than in the FOLFOX4 regimen group(P < 0.05). Conclusion: DOF regimen is effective in treating advanced gastric cancer and the toxicities can be tolerated. It is worth clinical application and extension.