This study aims to investigate the impact of the Notch signaling pathway on the invasion of Ameloblastoma (AM) and its influence on the expression of cyclinD1 protein. AM cells and dental follicle (DF) cells were cultured in vitro. The proliferation activity of two cells were measured using the CCK8. The invasive ability of cells were assessed using the Transwell chamber. A comparison was made between the proliferation and invasion abilities of two cells. The protein expression of cyclinD1, E-cadherin, and Snail was detected using Western Blot. The Notch signaling pathway was blocked using the Notch inhibitor FLI-06.The comparison of proliferation and invasion abilities between two cells were higher than control group cells.The difference was statistically significant (P<0.05). The Notch inhibitor on the invasive ability of AM cells is significantly lower than control group. The difference was statistically significant (P<0.05). After intervention with FLI-06, the protein expression of CDK 1, cyclinD1, and Snail in AM cells showed a significant downregulation, while the protein expression of E-cadherin showed a significant upregulation.The Notch signaling pathway plays a crucial role Notch signaling pathway promotes invasion ofAM by regulating E-cadherin via cyclinD1.