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Objective: The monosaccharide composition of Poria cocos polysaccharide (PCOS) from Poria cocos residue extracted by hot water, as well as its anti-cancer, anti-inflammatory factors and immune activities were studied, providing a theoretical foundation for the development of PCOS related products. Methods: The monosaccharide composition of water-extracted PCOS was analyzed by HPLC and FTIR, and the anti-tumor and immune activities of water-extracted PCOS were studied by CCK-8 method. Results: The polysaccharide content of PCOS was 76%, mainly composed of glucose, mannose, galactose and fucose, and the molar ratio was 1:0.107:0.079:0.018, with obvious polysaccharide characteristic absorption peak. The proliferation of gastric cancer cells, breast cancer cells, and liver cancer cells was decreased, with the best inhibition impact on gastric cancer cells (IC50=1096 µg/mL), and spleen lymphocyte proliferation in mice was enhanced. Conclusion: The water-extracted PCOS has good anti-cancer and immune activities in vitro.