Objective: To determine the sensitivity of different differentiated gastric cancer cell lines to various chemotherapeutic agents. Methods: The poor-differentiated gastric cancer MGC-803 cells and well-differentiated gastric cancer AGS cells as well as the normal gastric epithelial GES-1 cells were used, which were exposed to different concentrations of 5-fluorouracil (5-FU), oxaliplatin (L-OHP), irinotecan (CPT-11), docetaxel (DXT) and cisplatin (DDP) for 48 h respectively; the inhibitory effects of the chemotherapy drugs on those cells were determined by MTT assay, and the half maximal inhibitory concentrations (IC50) of the drugs were also calculated. The MGC-803 and AGS cells were exposed to IC50 concentrations of 5-FU, DDP and L-OHP for 48 h, respectively, and then the cell apoptosis and cell cycle distribution post treatment were measured. Results: All the 5 drugs exerted inhibitory effects on MGC-803, AGS and GES-1 cells (all P<0.05), in which, the effects of 5-FU, DDP and CPT-11 were relatively strong against AGS cells, L-OHP and DXT were relatively strong against MGC-803 cells, while DDP was relatively strong against GES-1 cells. Apoptosis was significantly increased in both types of gastric cancer cells after either of 5-FU, DDP or L-OHP exposure, and the apoptosis rate in 5-FU or L-OHP treated group of cells was higher than that in DDP treated group of cells (both P<0.05). Cell cycle analysis showed that L-OHP augmented S-phase arrest in both types of cells, DDP augmented S-phase arrest in MGC-803cells, and 5-FU and DDP augmented G1-phase arrest in AGS cells (all P<0.05), whereas 5-FU exhibited no obvious effect on the cell cycle distribution of MGC-803 cells (P>0.05). Conclusion: The inhibitory effects of chemotherapy drugs against gastric cancer cells are associated with the pathological types; and their effects on induction of apoptosis and the arrest of the cell cycle also varies among different gastric cancer cells.