Breast cancer is second most common in women and accounts for 23% of all occurring cancers in women. At present, more than 60% of the chemotherapeutic drugs are developed from plants and their derivatives, which can be used for development of anticancer drugs. An Indian barberry, Berberis aristata has been traditionally used for the treatment of inflammation, skin diseases, ulcers and cancers. In present study, the methanolic extract of stems of B. aristata, was used to investigate its anticancer activity in human breast cancer cell line (MCF-7). Different concentrations of the methanolic extracts (125, 250 and 500 μg/ml) were subjected to determine the cytotoxic effect by measuring the cell proliferation activity in MCF-7 breast cancer cell lines up to 48 h of incubation. The IC50 value for methanolic extracts identified was 220μg. Further, significant decreased (80%: p≤0.001) colony formation at 500μg/ml of methanolic extracts was noticed by soft agar assay in MCF-7 cells. However, in vitro scratch assay revealed the significant (p≤0.001) inhibition of cell migration up to 50% at 250μg of extracts. In addition, significant (68%) increase of apoptosis at 500μg of extracts in MCF-7 cells was evidenced by live/dead assay.