In this study, we examined the effect of acacetin on the apoptosis induction of HeLa human cervical cancer cells. The results showed that acacetin inhibited the cell viability and induced apoptosis, leading to PARP cleavage and activation of caspase-9, -3, and -7. Moreover, acacetin-induced apoptosis was attenuated by a broad caspase inhibitor, z-VADfmk. Also, acacetin resulted in a loss of mitochondria membrane potential. Taken together, our results demonstrate that the induction of apoptosis by acacetin in HeLa cells is associated with caspase activation via the mitochondria pathway.