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The aim of this study was to analyze the preventive effect of water-soluble polysaccharides in the maitake mushrooms (Grifola frondosa) fruiting body (GFP) on colon cancer in high-fat (HF) mice and WIDR cells. In vivo, the administration of GFP decreased colon myeloperoxidase (MPO) activity and disease activity index (DAI) compared with HF group (P < 0.05), and GFP was significantly inhibited the nitric oxide (NO) while increased of L-Glutathione (GSH) and superoxide dismutase (SOD). The expression of COX-2、IL-6、TNF-α was decreased in GFP -treated group, and the expression of lnterleukin-10(IL-10) and lnterleukin-4(IL-4) was increased. In addition, GFP can regulate the Wnt/β-catenin/GSK-3β signaling pathway to achieve the inhibitory effect on the survival of colon cancer. And CPS-II had high selectivity to tumor cells in vitro. The results revealed that CPS-II can inhibit colon cancer growth in vitro and in vivo, and may be a novel therapeutic agent for the disease.