AIM: To explore the effect and mechanism of AMI-1 on the proliferation, migration and apoptosis of human pancreatic cancer MIAPaca-2 cells. METHODS: MIAPaca-2 cells were randomly divided into the control groups and the treatment groups with different concentrations of AMI-1 (0.6, 1.2 and 2.4 mmol/L). MTT assay, colony formation assay, scratch assay and Transwell assay respectively were used to detect the effects of AMI-1 on the proliferation, colony formation and migration of MIAPaca-2 cells. The cell apoptosis was detected by flow cytometry. The effect of AMI-1 on the protein expression of caspase3, cleaved-caspase3, PRMT5, H4R3me2s and PCNA in MIAPaca-2 cells were detected by western blot. RESULTS: Compared with control group, after treatment with different concentrations of AMI-1, the cell survival rate of MIAPaca-2 cells decreased gradually in a time- and dose-dependent manner (P<0.01), the rate of cell clone formation decreased (P<0.01), the cell migration ability were diminished (P<0.01), the apoptosis rate was increased (P<0.01), and the protein level of cleaved-caspase3/caspase3 were increased, the protein level of PRMT5, H4R3me2s and PCNA were reduced (P<0.01). CONCLUSION: AMI-1 significantly inhibited the proliferation migration and apoptosis of pancreatic cancer cells in vitro, which may be associated with AMI-1 down-regulating the expression of PRMT5, H4R3me2s and PCNA, and up-regulation the expression of cleaved-caspase3/caspase3.