Ursolic acid, extracted from the traditional Chinese medicine bearberry, can induce apoptosis of gastriccancer cells. However, its pro-apoptotic mechanism still needs further investigation. More and more evidencedemonstrates that mitochondrial translocation of cofilin-1 appears necessary for the regulation of apoptosis.Here, we report that ursolic acid (UA) potently induces the apoptosis of gastric cancer SGC-7901 cells. Furthermechanistic studies revealed that the ROCK1/PTEN signaling pathway plays a critical role in UA-mediatedmitochondrial translocation of cofilin-1 and apoptosis. These findings imply that induction of apoptosis by ursolicacid stems primarily from the activation of ROCK1 and PTEN, resulting in the translocation of cofilin-1 fromcytoplasm to mitochondria, release of cytochrome c, activation of caspase-3 and caspase-9, and finally inducingapoptosis of gastric cancer SGC-7901 cells.