Objective To investigate the effects and mechanism of deoxypodophyllotoxin on cell proliferation and mi?gration of human lung cancer NCI-H358 cells in vitro. Methods CCK-8 assay, flow cytometry assay, wound healing assay and DCFH-DA assay were used to detect the effects of deoxypodophyllotoxin on the proliferation, cells cycle, apoptosis, mi?gration and reactive oxygen species (ROS). The protein expressions of Cyclin B1, Cdc25c, CDK1, Caspase-3, p53, Bcl-2, MMP9, ERK1/2, p38MAPK and JNK were measured by Western blot assay, respectively. Results Deoxypodophyllotoxin inhibited cell proliferation and reduced migration in human lung cancer NCI-H358 cells. Flow cytometry analysis showed that treatment with deoxypodophyllotoxin resulted in cell cycle G2/M and S phase arrest, cell apoptosis and ROS production. The result of Western blot assay showed that protein expressions of Cyclin B1, Cdc25c, CDK1, Bcl-2 and MMP9 were down-regulated while Caspase-3 and p53 were up-regulated. Moreover, Deoxypodophyllotoxin treatment decreased the phosphory?lated levels of ERK1/2, p38MAPK and JNK obviously. Conclusion Deoxypodophyllotoxin could suppress the proliferation and migration of human lung cancer NCI-H358 cells in vitro, which is a potential anti-tumor drug.