Objective: To investigate the effect of sodium citrate (SCT) on promoting apoptosis in gastric cancer cells and its mechanism. Methods: Gastric cancer MGC-803 cells were exposed to SCT (5, 10, 20 mmol/L) or 5-FU (0.5 mmol/L) respectively, using untreated MGC-803 cells as negative control. Then, the cell cycle distribution and apoptotic rate were analyzed by flow cytometry, the intracellular lactate content, phosphofructokinase-1 (PFK-1) activity and adenosine triphosphate (ATP) level were examined by colorimetric assay, and the expression of Bcl-2, Bax, caspase-3 and Cyt-c in MGC-803 cells were determined by Western blot analysis. Results: In SCT treated MGC-803 cells compared with negative control cells, the apoptosis and G2/M arrest were significantly increased, the intracellular lactate content, PFK-1 activity and ATP level were significantly reduced and Bcl-2 expression was significantly down-regulated, while the expressions of Bax, caspase-3 and Cyt-c were significantly up-regulated (all P<0.05). 5-FU exerted no significant effect on lactate content and PFK-1 activity in MGC-803 cells (all P>0.05), but all other effects were similar to those of SCT. Conclusion: SCT can promote apoptosis in MGC-803 cells, and the mechanism may be associated with its reducing glycolysis via inhibiting PFK-1 activity and activating mitochondria-dependent apoptotic pathway.