Semecarpus anacardium Linn. is a commonly used Ayurvedic medicinal plant which nuts have been described in Ayurveda and Sidda system of medicine to treat clinical ailments such as vitiligo, inflamation, microbial infection, geriatric problem, baldness and neuro related problems. In this study, anti-cancer activity of the leaves of Semecarpus anacardium Linn was evaluated for future drug development. The phytochemical screening was done by GC-MS analysis, cytotoxicity was examined using MTT assay, mode of cell death was evaluated by fluorescence microscopy and finally antitumor activity was determined in EAC cell induced tumor bearing mice. The ethyl acetate extract from the leaves of the plant induced cytotoxicity in cancer cells in a dose dependent manner (IC50: 0.57 µg/ml in MCF-7 cells) in different cancer cell lines. The non-malignant cells were relatively insensitive to the extract. The staining with acridine orange, ethidium bromide and DAPI confirmed that the extract induced apoptosis in cancer cells. Furthermore, the extract induced cell cycle arrest at G1 phase and suppressed cancer cell migration. An oral administration of the extract suppressed the tumor growth in mice model bearing ehrlich ascites carcinoma cells. The ethyl acetate extract was also found to prolong the survival of tumor bearing mice. Overall, these observations suggest the anticancer activities of the ethyl acetate extract of the leaves of S. anacardium. The study opens a new window to examine the phytochemical constituents from the leaves of the plant responsible for the anticancer activities.