Objective To investigate the effect of Bortezomib on cell cycle, proliferation, apoptosis and nuclear factor-κB (NF-κB) in cells. Methods Methyl thiazol tetrazolium (MTT) method was used to detect the inhibitory effect of Bortezomib on cell growth, analyze the effect of Bortezomib on cell cycle and apoptosis by flow cytometry. The effect of Western blotting on NF-κB, inhibitor of NF-κB (IκB) and Bortezomib to detect the expression of B cell lymphoma/leukemia-2 (bcl-2). Results (1) The inhibitory effect of Bortezomib on gastric cancer cells was dose-dependent and inhibited with increasing concentration (t=7.418, 5.131 and 6.438, P=0.001, 0.001 and 0.005). (2) 95D cell apoptosis peak appeared, and the control group accounted for the proportion of G2/M phase cells were (36.97±11.46)% and (19.17±5.37)%, respectively. The difference was statistically significant (P=0.031); 973 cell apoptosis peak, compared with the control group, G2/M phase cells accounted for the proportion were (40.34±13.83)% and (13.10±4.72)%, respectively. The difference was statistically significant (P=0.037); GLC82 cells neither apoptosis peak appeared, no obvious changes in cell cycle, the cell ratio of G2/M and the control group were (20.37±6.14)% and (12.15± 2.63)%, respectively. Two groups had no statistically significant difference (P=0.173). (3) Basic expression of NF-κB was found in gastric cancer cells, and the expression level of NF-κB in nucleus was inversely proportional to the expression level of IκB. No significant effect on the expression level. (4) Bortezomib could significantly regulate the level of anti apoptotic protein bcl-2, and the inhibition was in a time and dose-dependent manner. After different concentration of Bortezomib, the expression level of NF-κB in nucleus decreased in different degrees, the difference was statistically significant (t=6.373, 4.273, 3.841 and 4.414, P=0.006, 0.018, 0.026 and 0.015). Conclusion Bortezomib can inhibit the proliferation of gastric cancer cell line and induce apoptosis, which may play a role in NF-κB pathway. Key words: Gastric cancer; Bortezomib; Nuclear factor-κB; B cell lymphoma/leukemia-2