Objective To investigate the influence of CO2-insufflation pressure on invasion potential of the colon cancer cells. Methods With an in vitro artificial pneumoperitoneum model, SW1116 human colon cancer cells were exposed to CO2-insufflation of 5 different pressure groups: 6, 9, 12, 15 mm Hg and control group, respectively for 1 h. The invasion capacities of SW1116 cells exposed to CO2-insufflation of 5 different pressure groups were detected by cell adhesion/invasion assay in vitro. Results Immediately following exposure to 15 mm Hg CO2 insufflation, the invasion of SW1116 cells decreased significantly compared to the cells before exposure. At the 0 h time point, the cells exposed to 15 mm Hg were significantly less invasive than those exposed to the other insufflation pressure (P＜0.05), and the cells exposed to 6 mm Hg were more invasive than cells exposed to the other insufflation pressure (P＜0.05). And 72 h after exposed to CO2-insufflation, the differences between the pressure groups were not significant. Conclusion CO2-insufflation induced a temporary change in the invasion capacity of cancer cells in vitro, higher pressure of CO2-insufflation inhibits the invasion potential.