The objective of this study was to examine the cytotoxic effect of BarleriaacuminataNees plant in rootand stem extract on cervical cancer (HeLa) cell lines. HeLa cells were cultured in EMEM medium and incubated with different concentrations (12.5, 25, 50, 100 and 200 µg/ml) of ethanol extract of B. acuminata. The method used for this research to determine cell viability wasan 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.B. acuminate decreased cell viability in malignant cells in a concentration dependent manner. The IC50 value of B. acuminataextract was found to beroot 166.1µg/mland stem168.2 µg/ml. These data showed ethanolic extract of B. acuminatareduces the viability of cervical cell lines and may have potential as anti-cancer compounds.