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Cancer is a frightful disease and represents one of the biggest health-care issues for the human race and demands a proactive strategy for cure. Exploration of natural product containing anticancer agent provide a promising line for research on cancer. Moringa plant (Moringa oleifera) is one of the medicinal plants used in traditional medicine for the treatment of cancer. Several studies have reported that water and alcoholic leaves extracts of M. oleifera have anticancer activity in some cancer cell line, including HepG2 liver cancer cells, A549 lung cancer cells, Caco-2 colon cancer cells and MDA-MB-231 breast cancer cells, but study on T47D breast cancer cells has not reported yet. Therefore, this study was carried out to investigate the cytotoxic activity of ethyl acetat fraction of M. oleifera (EMO) on T47D cells by observing cell cycle profile. Cytotoxic activity was determined by MTT assay. Cell cycle analysis was performed using flow cytometry and cyclin D1 expression was analyzed using immunocytochemistry method. Based on preliminary MTT assay, T47D treated with EMO demonstrated a medium cytotoxic effect with an IC50 value of 135.312 ug/mL. Further assay indicated that EMO induced cell cycle arrest on G0/G1 phases. Immunocytochemistry assay showed that the EMO decreased expression of cyclin D1.