Chitosan oligosaccharide (COS) represents a class of natural polymers that holds therapeutic promise in several diseases due to not only its physiochemical properties including water-solubility and low viscosity but also its favorable pharmacological properties including good pharmacokinetics and safety profiles and diverse beneficial biological activities. Activation of AMPK and inhibition of inflammatory signaling pathways including NF-κB and MAPK pathways are recognized as major mechanisms responsible for several effects of Chitosan oligosaccharide (COS) including anti-inflammation, anti-cancer, and anti-diabetes. COS can interrupt cancer progression at multiple stages by modulating several signaling proteins/pathways including NF-κB, AMPK, mTOR, caspase-3, CD147, MMP-2, MMP-9, and VEGF. In vitro experiments have demonstrated that Chitosan oligosaccharide (COS) induced the death of several cancer cell types including ascites, bladder cancer, prostate cancer, lung cancer, liver cancer, leukemia, cervical cancer and colorectal cancer. The values of IC50 of Chitosan oligosaccharide (COS) in inducing cytotoxicity are 25 μg/mL-50 μg/mL depending on types of cancer cells.