Objective To study effect of bufalin on invasion and metastasis of gastric cancer cells and related mechanism.Methods AGS human gastric cancer cell line was used for in vitro experiments.Cultured cells were treated by negative control group,bufalin group and oxaliplatin group.Cell proliferation was determined by MTT.Invasion and metastasis were observed by Wound-Healing Assay and Transwell Assay.Expression levels of E-cadherin,matrix metalloproteinases (MMP)-2,MMP-9 were detected by Western blotting.Results Bufalin was found to significantly inhibit the proliferation of AGS cells in a dose and time dependent manner.Wound-Healing Assay and Transwell Assay showed that as compared with the blank control group,bufalin inhibited invasion and metastasis of AGS cells (P＜ 0.05),but there was no statistically significant difference between bufalin group and oxaliplatin group (P＞0.05).Western blotting showed the expression of E-cadherin was increased in bufalin group as compared with the blank control group while the expression levels of MMP-9 and MMP-2 were down-regulated (P＜0.05),but there was no statistically significant difference between bufalin group and oxaliplatin group (P＞ 0.05).Conclusion Bufalin has anti-cancer activity on gastric cancer cells,and it has the ability of inhibiting cancer cell invasion and metastasis,and regulating the expression of some related gene.