Objective:To evaluate the effect of pristimerin on the migration and invasion of non-small cell lung cancer(NSCLC)NCI-H1299 cells,and explore the underlying mechanism. MethodsNCI-H1299 cells were treated with different concentrations of pristimerin (0~80 μmol/L). The proliferation of NCI-H1299 cells was detected by cell counting kit (CCK-8) assay, and the non-toxic dose of pristimerin to NCI-H1299 cells was determined, which was used as the follow-up dose for the study of pharmacodynamics and molecular mechanism. The effect of pristimerin on the lateral migration ability of NCI-H1299 cells was evaluated by wound-healing assay; the effect of pristimerin on the longitudinal migration and invasion ability of NCI-H1299 cells was evaluated by transwell migration and invasion assay; the protein and mRNA expression levels of epithelial-mesenchymal related markers were measured by Western blot and real-time quantification PCR assay, respectively.  Results:When the concentration of pristimerin was less than 2.5 μmol/L, it had no obvious cytotoxicity to NCI-H1299 cells. Compared with the blank control group, the non-toxic dose of pristimerin (0.5 μmol/L, 1.0 μmol/L and 2.0 μmol/L) could significantly inhibit the lateral migration and longitudinal migration and invasion of NCI-H1299 cells (P<0.01). After treatment with pristimerin, the expression levels of epithelial markers E-cadherin and ZO1 were significantly increased, while the expression levels of mesenchymal cell markers N-caherin and Vimentin were significantly decreased (P< 0.01).  Conclusion: Pristimerin can inhibit the migration and invasion of NCI-H1299 cells, and this effect may be related to the regulation of epithelial-mesenchymal transformation of NCI-H1299 cells.