Objective: To investigate the role of glycogen synthase kinase 3β (GSK-3β) in cadherin 17 (CDH17)-mediated of gastric cancer cell invasion and the mechanism. Methods: Gastric cancer MKN-45 cells were transfected with CDH17 siRNA or exposed to GSK-3β inhibitor SB216763 respectively, with untreated and empty vector transfected MKN-45 cells as blank control and negative control, respectively. The changes in GSK-3β, β-cantenin and NF-κB (p50/p65) expressions and invasion ability of each group of cells were determined. Results: Compared with blank control cells, in MKN-45 cells transfected with CDH17 siRNA, the CDH17, phosphorylated GSK-3β, β-cantenin and p50/p65 expressions were significantly decreased, and the number of invaded cells was significantly reduced (all P<0.05). In MKN-45 cells exposed to SB216763, the CDH17 and β-cantenin expressions showed no significant difference (both P>0.05), while the phosphorylated GSK-3β and p50/p65 expressions were significantly decreased, and number of invaded cells was significantly reduced (all P<0.05). In MKN-45 cells transfected with empty vectors, all parameters showed no significant change (all P>0.05). Conclusion: In the signaling pathway of CDH17-medated gastric cancer cell invasion, GSK-3β may be an important molecule downstream to β-cantenin for regulating NF-κB activity through its phosphorylation level.