The main objective of the present research work was the isolation of the bioactive molecule vitexin present in the methanolic extract of beet root (Beta vulgaris) and evaluation of antitumor activity. Vitexin was characterized by HPLC-MS, IR, H1-NMR, 13C-NMR etc. Molecular docking of vitexin with respect to the target protein Topoisomerase-I was evaluated by Auto dock program with PDB id 1A36 and displayed the binding energy -3.95 k.cal/mol. In vivo antitumor activity of ME-BRT was carried out against S-180-AC rat model. ME-BRT (100 mg/kg), ME-BRT (200 mg/kg) significantly increased the PILS. While topotican increased the life span of S-180-AC 78.57%, ME-BRT (100 mg/kg) increased it by 57.14% and 71.42 % ME-BRT (200 mg/kg) respectively. So ME-BRT at dose 100 and 200 mg/kg significantly improved the overall survival of all treated animals and topotecan was not significantly differed from each other in improving the overall survival of S-180-AC. Both extracts displayed a significant inhibition of tumour growth at doses of 100 and 200 mg/kg with values of 81.90 and 88.33%, respectively, with no mortality, compared standard drug topotecan (95.94%).