Bcl2 family proteins Bcl-2 and Bcl-xL play an important role in the inhibition of apoptosis and autophagy, inhibitors of these proteins are potential compounds functioning in cancer therapy. Here we identified a novel Bcl-2/Bcl-xL inhibitor Z40 via a combination of structure-based design, surfase plasmon resonance method and NMR-based structural analysis. Z40 efficiently induced cell death and inhibts cell proliferation in Hela cells.Detailed analysis showed that Z40 induces chromatin compaction, DNA condensation and DNA fragmentation. According to the results of Hoechst33342/PI staining cooperating with flow cytometry analysis, portion of the Z40-treated Hela cells displayed high blue and low red fluorescence. These results provided evidences that Z40 induced apoptotic phenomenon in Hela cells. Z40 also induced cytochrome c release and Activation of caspase-3. Hela cells pretreaded with zDEVD-fmk showed decreased cell death induced by Z40, indicating that Z40 induced caspase-3-dependent cell death.Further studys revealed that Z40 induced punctate distribution of GFP-LC3, accumulation of GFP-LC3-II and LC3-II, formation of double-membraned autophagosomes and cell death inhibited by the presentation of 3-MA, indicating that Z40 also induced autophagic cell death in Hela cells.All togrther, these reasults showed that inhibitor of Bcl-2/Bcl-xL could induce both autophagic cell death and apoptosis in cancer cells.