Malignant gliomas are the primary malignant brain tumors which are difficult to remove completely by surgery. Adjuvant chemotherapy turns out to be one of the alternative treatments. Autophagy, a type II cell death, has been observed as a highly regulated process of degradation and reusing of intracellular proteins and organelles. Accumulated evidence demonstrated that various anticancer therapies induced autophagy among indicated cancer cell types. Evodiamine, one of the major bioactive compounds isolated and purified from Chinese herbs, Wu-Chu-Yu, has the ability to induce apoptosis in different types of cancers. However, the signaling mechanism of evodiamine-induced autophagy remains elusive. For provide a strategy for development of new chemotherapeutic agents, the specific aim of this study is to investigate the molecular signaling of evodiamine-induced cell death in U87-MG, a human malignant glioma cell line. We performed MTT assay to examine the effect of evodiamine on the growth ability of U87-MG cells. Results showed that evodiamine significantly inhibited the growth of U87-MG cells (p<0.001). As revealed by flow cytometry, we demonstrated that the percentage of autophagy reached a plateau of 42.3 ± 7.5 % (p<0.001) after 24 h exposure of 6 μM evodiamine. Using immunoblot assay, we also detected the processing of microtubule-associated protein 1 light chain 3 (LC3), indicating that evodiamine is able to induce autophagy in U87MG cells. A parallel experiment also indicated an apoptosis phenomenon which reached to 33.0 ± 3.9 % (p<0.001) after 48 h exposure of 6 μM evodiamine, by using Annexin V/Propidium iodide (PI) double staining . These results indicated that autophagy might play as a prelude to apoptosis in evodiamine-treated U87-MG cells. Moreover, a rapid and marked increasement of intracellular calcium (〔Ca2+〕i) was detected 1 h after evodiamine-treated U87-MG cells . Pre-treatment of an endoplasmic reticulum IP3 receptor blocker, 2-APB, could prevent both intracellular calcium (38%) and evodiamine -induced autophagy (45%). Using JC-1 staining, we also detected that mitochondrial membrane depolarization . A MPTP blocker, cyclosporine A (CsA) decreased the evodiamine-induced autophagy and increased evodiamine-induced apoptosis, suggesting that evodiamine-induced autophagy was regulated by mitochondrial damage. In conclusion, our results indicated that evodiamine induce both autophagy and apoptosis in U87-MG cells, however, autophagy may ahead from and inhibit the apoptosis and these phenomena are involved with mitochondria membrane depolarization and intracellular calcium.