Two new acetophenone derivatives were isolated from the roots of Euphorbia ebracteolata Hayata, their structures were elucidated as 2,4-dihydroxy-6-methoxyl-methylene-3-methyl-acetophenone (1) and 2-dihydroxy-6-methoxyl-methylene-3-methyl-acetophenone-4-O-beta-D-glucopyranoside (2) on the basis of chemical and spectroscopic methods including (1)D- and (2)D-NMR techniques, and their cytotoxic activities were investigated by using the MTT method.