Gastric cancer is a common gastrointestinal malignancy that accounts for a notable proportion of cancer-associated mortalities worldwide. Cytoplasmic fragile X mental retardation 1-interacting protein 2 (CYFIP2) is a novel p53-mediated pro-apoptotic protein whose expression is decreased in gastric cancer. However， whether decreased expression of CYFIP2 contributes to gastric carcinogenesis remains unclear. In order to mimic gastric tumor CYFIP2 expression levels， the present study used short hairpin RNA targeting CYFIP2 mRNA to silence CYFIP2 expression in MGC803 and SGC7901 gastric cancer cells. Gastric cancer cells with constitutively decreased CYFIP2 expression levels were successfully established. It was observed that CYFIP2 knockdown promoted proliferation and colony formation， and inhibited apoptosis in these cells. Furthermore， 5-fluorouracil (5-FU)-induced apoptosis was decreased following inhibition of CYFIP2 expression. In SGC7901 cells， protein expression of active caspase-3 and cleaved poly (ADP-ribose) polymerase was increased following treatment with 5-FU， while phosphorylated Akt serine/threonine kinase 1 (Akt) levels were decreased. These 5-FU-induced effects were reduced following CYFIP2 knockdown. In addition， inhibition of the Akt signaling pathway using the Akt inhibitor LY294002 restored CYFIP2-knockdown SGC7901 cell chemosensitivity to 5-FU. The results of the present study demonstrate that decreased CYFIP2 expression is associated with increased gastric tumor growth and that CYFIP2 knockdown-induced activation of the Akt pro-survival signaling pathway confers resistance to 5-FU-based chemotherapy in gastric cancer cells. Therefore， combined treatment with an Akt inhibitor and chemotherapeutic drugs may improve the efficacy of gastric cancer therapy in patients with low CYFIP2 expression.