The pathogenesis of gastric cancer is characterized by excessive proliferation, abnormal differentiation, and reduced apoptosis. Ursolic acid, extracted from traditional Chinese medicine bearberry, inhibits cell growth and induces apoptosis in gastric cancer. However, the mechanism of the proapoptotic effect of ursolic acid on gastric cancer cells needs further investigation. In our present study, we found in apoptotic gastric cancer BGC-823 cells induced by ursolic acid that a translocation of cofilin-1 protein from the cytoplasm to the mitochondria promoted the release of cytochrome c from the mitochondria to the cytoplasm, thereby activating the caspase cascade and finally inducing gastric cancer cell apoptosis. These results implied that the mitochondrial translocation of cofilin-1 might play a crucial role in the promotion of apoptosis and might be a key target for future treatment of human gastric cancer.