Scolopendra subspinipes mutilans, also known as Chinese red-headed centipede, is a venomous centipede from East Asia and Australasia. In traditional Chinese medicine, this centipede has been widely used for the treatment of various ailments, but few researches on the venom from this centipede have been conducted so far. In the present study, we reported that the venom could potently suppress the growth of the myelogenous leukemia cell lines K562, U937 and HL- 60, in a dose-dependent manner with IC50 values of 20 μg/mL, 30 μg/mL, 200 μg/mL, respectively. K562 cells treated with the venom showed typical morphological indicators of apoptosis including condensation of nuclei. Annexin V-FITC and propidium iodide double staining further demonstrated that the venom had potent apoptogenic activity. Venom treatment induced caspase 2, 3, 6, 8 and 9 activation in K562 cells, which might involve in the venom-induced apoptosis. The venom could also cause cell cycle arrest. Increased cell accumulation at S phase was observed in K562 cells treated with the venom. Taken together, the Chinese red-headed centipede venom could inhibit the growth of myelogenous leukemia cell lines through apoptosis induction and cell cycle arrest. It might be a useful source for the identification of substances with anti-leukemia activity.