Previous studies have shown that the Ginkgo biloba extract (EGb761) can be used to anti-cancer. However, the mechanism by which EGb761 mediate this effect is still unclear. In the present study, EGb761 inhibited cell proliferation and induced cell apoptosis in gastric cancer cell was explored. The cell viability was detected by the CCK8 assay. The cell cycle and apoptosis was assessed by flow cytometry. The protein expression of caspase-3, p53 and Bcl-2 were analyzed by western blot. Treatment of human gastric cancer cells with EGb761 induced cell death in a dose-dependent manner by using CCK8 assay. Consistent with the CCK8 assay, the flow cytometry results showed that gastric cancer cells were accumulated in G0/G1 phase when exposed to EGb761. Furthermore, the proportion of apoptosis cells was increased after EGb761 treatment as compared to untreated group. In addition, our results showed that the treatment of AGS cells with EGb761 significantly increased the expression of caspase3 and p53, and decreased the anti-apoptotic Bcl-2 level. Our results demonstrated that EGb761 could inhibit gastric cancer proliferation through adjusting cell cycle and inducing cell apoptosis.