Abstract: The polyphenols were extracted from the Leccinellum crocipodium by ethanol as extraction solvent extraction and Folin-Ciocalteu method. The effect of ethanol concentration,temperature,solid-liquid ratio and time on the extraction efficiency of polyphenols were identified. Based on single factor test,an orthogonal design was applied to optimize extraction process. The polyphenol composition and inhibitory effect on Caco-2 cells of Leccinellum crocipodium were investigated in the present work. The results showed that the optimum conditions for extraction of polyphenols were temperature of 80 ℃,ethanol concentration of 50%,solid-liquid ratio of 1∶25,extracted 1 h,two times. Under this condition,polyphenol extraction rate was 5.46% ±0.04%.Among 18 kinds of polyphenols,the polyphenols of this wild edible fungi were mainly composed of 5-sulfosalicylic acid,catechin,cinnamon and other five kinds of polyphenols,and the content of 5-sulfosalicylic acid up to779.00 mg/100 g. The inhibitory effects on colon carcinoma cells showed that the cell inhibition rate was up to55.07%,when the polyphenols concentration was 175 μg/m L.