Using plant-based treatment approaches and herbal medicines is getting attention these days to avoid the side effects of the current treatment methods for cancer. According to the literature, bitter melon extract exhibits cytotoxic effects against cancer by affecting the hormonal pathways. Therefore, in this research, the effects of the bitter melon extract with its combination of different concentrations of drugs were investigated against different cell lines (MCF-7, MDA-MB-231, RAW 264.7, J774). Both hormone-positive and negative breast cancer cell lines were used and the obtained data were compared with the literature. As a result, when exposed to bitter melon extract, the hormone-positive breast cancer cell line MCF-7 exhibits more cytotoxic effects compared to the triple hormone-negative breast cancer cell line MDA-MBA-231. For the combination of drugs along with the bitter melon extract; the highest cytotoxicity was observed with Tamoxifen.