Chitosan oligosaccharide (COS, 3 kDa<MW<5 kDa) was tested for colon cancer chemoprevention by measuring the activities of quinine reductase (QR) and glutathione-S-transferase (GST), glutathione (GSH) levels, ornithine decarboxylase (ODC) activity, and cyclooxygenase (COX)-2 expression in HT-29 cells treated with COS. COS induced QR activity in a dose-dependent manner over a concentration range of 0.1-4.0 mg/ml. GST activity was also induced in HT-29 cells treated with COS. In addition, GSH levels were increased 1.3-, 1.4-, and 1.5-fold with COS at 2, 3, and 4 mg/ml, respectively. ODC activity induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) was inhibited by 33% and 39% with 3 and 4 mg/ml of COS, respectively. COS also inhibited the expression of TPA-induced COX-2 protein in HT-29 cells. These results suggest that COS has colon cancer chemopreventive activity by increasing QR and GST activities and GSH levels and by inhibiting ODC activity and COX-2 expression in vitro.