Beta Vulgaris Linnaeus ranks among the 10 most powerful vegetables as excellent sources of phytochemicals, which showed potent antioxidant and anticancer activities. Aim of study: To determine the cytotoxicity and activity of B. Vulgaris root extracts against colon carcinoma cells. Methods: The Beetroot extracted into ethanol/water (80/20 v/v) solvent at a solid/liquid ratio of 1/3 (g/mL) for 1 hr, HCT-116 cells (human colon carcinoma cell line) were obtained from VACSERA Tissue Culture Unit. The effect of extract was performed using MTT assay and the evaluation of Apoptotic mechanism of cytotoxicity against HCT-116 cell line also Cell Cycle was analyzed using Flow Cytometry. The results: The MTT cytotoxicity assay revealed that the beet-root ethanol extract has an obvious cytotoxicity on HCT-116 cancer cell lines at low significant IC50 value as104±4. 7µg/ml. Inhibitory activity against colon carcinoma cells was detected under these experimental conditions in vitro. The total percentages of early, late apoptosis and Necrosis ratio after treatment with a beet-root extract and a control group (non-treated cells) for 48 hours in the HCT-116 cell lines were found to be 2.72%, 17.85%, 7.52%, 0.55%, 0.18% and 1.24% respectively. On these findings, the apoptosis-inducing effect of the beet-root extract is true. However, the apoptotic effects of beet-root extract on HCT-116 cell lines were significant. In conclusion: the present study indicates that treatment of cancer cell lines (Hct-116) with red beet-root extract inhibited the cell proliferation and significantly induced apoptosis at the level of the used extracts. However, the exact proliferative and apoptotic mechanisms of beet-root extract of different cancer cells is still unknown and further studies are needed in this regard.