The present study includes the preliminary phytochemical screening and evaluation of the anticancer potential of ethanolic as well as aqueous extract of whole plant Pajanelia longifolia (Willd) K.Schum. Anticancer activity of the whole plant were investigated by standard MTT assay method using Hacat and MCF-7 cell lines and the IC50 values were calculated. The invitro cytotoxic effect of ethanolic and aqueous extracts of the whole plant Pajanelia longifolia (Willd) K.Schum were identified. Among this ethanolic extract was found to be more effective against Hacat and aqueous extract possess more activity against MCF- 7. The present study concludes that both the extracts of whole plant of Pajanelia longifolia (Willd) K.Schum acquired anticancer activity against Hacat and MCF-7 cell lines.