Objective To investigate the effect and molecular mechanism of β-lapachone combined with NVP-BEZ235 on the proliferation and migration of BGC-823 human gastric cancer cells. Methods BGC-823 cells were randomly divided into four groups: control group, 1 μmol/L β-lapachone group, 50 nmol/L NVP-BEZ235 group and 1 μmol/L β-lapachone combined with 50 nmol/L NVP-BEZ235 group. The proliferation of cells was determined using the MTT assay and colony formation assay. The expression levels of proliferation-related proteins phosphorylated AKT (p-AKT), phosphorylated NF-κB (p-NF-κB), phosphorylated extracellular signal-regulated kinase (p-ERK) and cyclin D1 were detected by Western blotting. The migration of cells was measured by wound healing assay and TranswellTM migration assay. The expression levels of epithelial-mesenchymal transition (EMT) markers E-cadherin, vimentin, Snail and β-catenin were detected by Western blotting. Results Compared with β-lapachone or NVP-BEZ235 treatment, the combination of β-lapachone and NVP-BEZ235 showed more prominent inhibitory effect on the proliferation and colony formation of BGC-823 cells.The most effective suppression on the expressions of p-AKT, p-NF-κB, p-ERK and cyclin D1 was observed in the combination therapy. Combined treatment also showed more evident inhibitory effect on the migration of BGC-823 cells as compared with β-lapachone or NVP-BEZ235 treatment. The expression of E-cadherin was significantly up-regulated, but the expressions of vimentin, Snail and β-catenin were significantly down-regulated by the combined treatment. Conclusion β-lapachone combined with NVP-BEZ235 can effectively inhibit the proliferation of BGC-823 cells, which may be related to the down-regulation of p-AKT, p-NF-κB, p-ERK and cyclin D1. Moreover, the combined treatment can effectively suppress the migration of BGC-823 cells via modulating the EMT process.