Background: Carthami flos (CF) is a traditional medicine used to treat various diseases, especially cancer therapy. Objectives: The mechanisms of cell death induction by CF were investigated in AGS human gastric adenocarcinoma cells. Materials and Methods: Cell viability assay, cell cycle analysis, caspase activity assay, western blotting, and reactive oxygen species (ROS) assay were used to check the anti-cancer effects of CF on AGS cells. Results: Treatment with CF (100500 μg/mL) inhibited AGS cell proliferation and increased the ratio of the subG1 phase of the cell cycle. CF induced cell death was associated with a reduction in Bcl-2 and an increase in Bax levels. Moreover, expression of the apoptosismediating surface antigen (FAS) was increased. CF activated caspase-3 and -9. In addition, it regulated the activation of mitogen-activated protein kinases and increased intracellular ROS generation. Conclusion: These findings suggest that CF induced apoptosis in AGS cells and could thus, serve as a novel anticancer agent to promote apoptosis of gastric cancer cells.