The current research was designed to evaluate the phytochemical contents, cytotoxic and antimicrobial activity of Emex spinosa extracts. The different plant extracts and Aloe-emodin glucoside were screened using the colorimetric MTT method (3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl- tetrazolium bromide) assay to test their in vitro cytotoxic activity against HepG2, MCF-7, Caco-2 and HCT. The clinically used anticancer drug doxorubicin was used as standard for comparative purposes. Anthraquinones (Aloe-emodin-O-glucoside, Emodin and nataloin (1, 2, 8-trihydroxy, 6-methyl, 10-anthrone-C-glucoside) together with β-sistosterol and β-sitisterol-O-β-D-glucoside were isolated from Emex spinosa. Aloe-emodin glucoside together with four fractions from this plant were evaluated for their anticancer and antimicrobial activities. Aloe-emodin glucoside showed anticancer activity against HCT, HepG-2, MCF-7 and Caco-2 cell lines. The total ethanol extract of E. spinosa and diethyl ether, chloroform, ethyl acetate and butanol fractions shown antibacterial activity against Staphylococcus aureus, Streptococcus pyogenes and Bacillus subtilis.