Objective：To investigate the effect of tumor suppressor gene DLC-1 on Wnt/β-catenin signaling pathway in colon cancer cell line SW480. Methods：Human colon cancer cell line SW480 was taken as research object and was divied into 3 groups：pcDNA3.1（+）-DLC-1 group，pcDNA3.1（+） group and SW480 group. Each group was transfected with recombinant lentiviral vector containing DLC-1 gene. MTT was applied to detect cell proliferation and flow cytometry was performed to detect cell apoptosis. Real-time PCR and Western blot were preformed to detect the mRNA and protein expressions of Wnt/β-catenint signaling pathway related genes（β-catenin，GSK-3β and c-my）. Results：The proliferation of SW480 cells was significantly inhibited（the proliferation rate was 0.546）and the apoptosis was increased（the apoptosis rate was 42.422+3.413） in pcDNA3.1（+）-DLC-1 group. Real-time PCR and Western blot determined that the mRNA and protein expressions of β-catenin and c-myc were down-regulated，and the mRNA and protein expressions of GSK-3β were up-regulated（P=0.000）. Conclusion：DLC-1 plays a suppression role in colon cancer cell，which asso－ciates with Wnt/β-catenint signaling pathway. DLC-1 may be the upstream adjustment factor of Wnt/β-catenint signal pathway.