OBJECTIVE: In this study, we investigated the effects of ethanol extract of Hupiruisu Fang(HPRS), a formula of Traditional Chinese Medicine, on HCT116 cell line.METHODS: MKN45, A549, Hela and HCT116 cells were treated with ethanol extract of HPRS alone or the extract plus 5-Fluorouracil(5-FU) for 48 h, and then the cell viabilities were measured using CCK-8Kit. The early apoptosis rate and total apoptosis rate in both HCT116 cells were evaluated by flow cytometry. The m RNA levels of apoptosis-related genes including caspase-3, caspase-8, Bcl2 and Baxwere detected by real-time quantitative polymerase chain reaction. Lastly, the protein activities and expressions of those apoptosis related genes were observed for further verifying the pro-apoptosis of the extract of HPRS.RESULTS: Ethanol extract of HPRS could significantly induce apoptosis in HCT116 cell line. Synergistic analysis revealed that the extract exhibited a significant effect upon 5-FU-associated cytotoxicity in the cell line.CONCLUSION: The ethanol extract of HPRS plus5-FU might have the potential to improve the treatment of colorectal cancer.