Objective:To explore the effect of insulin on the pancreatic cancer cell lines and the potential mechanisms. Methods: The expression of insulin receptor(IR) was evaluated by qRT-PCR and Western blot. We examined the effect of insulin with different concentrations on the viability of pancreatic cancer cell lines to choose optimal concentration. After insulin treatment, the viability of pancreatic cancer cells was measured by CCK-8 assay; Invasion and migration of cells were measured by a Transwell chamber assay; Western blot was used to observe the expression of matrix metalloprotease(MMP)-2, MMP-7 and MMP-9. Results: Pancreatic cancer cell lines PANC-1 and Miapaca-2 showed relatively higher expression of IR(P<0.05). The cell lines had significantly enhanced viability(P<0.05) when treated with insulin of 100 nmol/L. CCK-8 assay showed that the proliferation of the cell lines was increased significantly. Transwell chamber assay showed that insulin significantly enhanced the invasion and migration of pancreatic cancer cells(P<0.05). Western blot proved that the expression of MMP-2, 7 and 9 was remarkably increased. Conclusion: Insulin can enhance the invasion and migration of pancreatic cancer cell lines by up-regulating MMP-2, 7 and 9.