The 70% ethanol extract of Anemone vitifolia in Enshi area of HuBei province was isolated and purified by column chromatography on silica gel, Sephadex LH-20 and preparative liquid chromatography. Structures were proved by physicochemical properties, NMR and MS.The antitumor activity was tested by MTT assay. Results 10 compounds, including lignin, flavonoidsand saponins, were obtained from 70% ethanol extract of Anemone vitifolia , and were identified trachelogenin (Ⅰ), arctigenin (Ⅱ), chlorogenic acid (Ⅲ), astragalin (Ⅳ), isoquercitrin (Ⅴ), tiliroside (Ⅵ), tracheloside (Ⅶ), arctiin (Ⅷ), salicin (Ⅸ) and dioscin (Ⅹ). CompoundsⅠ~Ⅱ、Ⅵ~Ⅶ、Ⅸ were isolated from the genus Amenone for the first time; Compounds Ⅰ~Ⅶ、Ⅸ、Ⅹ were isolated  from Amenone vitifolia for the first time. Compounds Ⅱ、Ⅷ、Ⅸ、Ⅹ inhibited the proliferation of HeLa cells with inhibition rates of 10.5%, 7.0%, 0.5% and 88.0% at 100 μmol/L, and the inhibition rate of compound Ⅹ was higher than that of the positive control cisplatin by 80.5%.