Anti-cancer effects of Nepeta Deflersiana Extract (NDE) in estrogen positive and negative forms of breast cancer

Aim: This study focused on efficacy of the ethanol extract gained from the aerial parts of Nepeta deflersiana (ND) on breast cancer (BC) cells. Materials and Methods: Shade-dried aerial parts of ND were extracted with ethanol using the Soxhlet extraction method (NDE). MTT assay was performed to assess the cytotoxicity of NDE in normal breast cells (MCF-10A), as well as estrogen receptor (ER) positive (MCF-7) and ER-negative (MDA-MB231) BC cells. Nuclear staining was performed using Hoechst/propidium iodide. Annexin V assays for apoptosis detection and cell cycle analysis were performed by flow cytometry. The effect of NDE on metastasis was analyzed by the transmigration assay. Western blot was used to detect the key signaling proteins. Results: NDE inhibited BC cell proliferation with GI₅₀ values of 154.90 and 202.7 μg/ml for MCF-7 and MDA-MB231 cells, respectively. NDE was non-toxic to normal MCF 10A cells at concentrations up to 1000 μg/ml. NDE induced nuclear condensation/fragmentation in both types of BC cells near the GI₅₀ doses values. An apparent increase in early and late apoptotic populations was confirmed by the flow cytometry. The extract also increased the sub-G₀/G₁ population in both types of BC cells. Dose-dependent increases in Caspase 3 and Bax levels were also recognized, as were NCE decreases in anti-apoptotic Bcl-2 levels in the same cell lines. Conclusion: NDE possessed excellent anticancer properties against ER-positive and negative BC cells, encouraging further research on the use of this natural product against BC.