Anticancer Activity of Miswak Root Extract in Breast Cancer Cell Line: HRLC-MS/MS Profiling, In Vitro Evaluation, and...

Breast cancer is among the most commonly diagnosed malignancies in women and remains difficult to treat due to therapy resistance and the adverse effects associated with conventional chemotherapeutic regimens. In this study, the anticancer activity of the ethanolic root extract of Salvadora persica (S. persica), commonly known as Miswak, was evaluated in human breast cancer cells using a combination of in vitro assays, phytochemical profiling, and computational analyses. HRLC-MS/MS characterization revealed a wide range of bioactive constituents, including alkaloids, flavonoid derivatives, glucosinolates, and fatty acid-based molecules detected under both ionization modes. The extract exhibited a concentration-dependent cytotoxic effect on breast cancer MCF-7 and MDA-MB-231 cells, with IC₅₀ values of 144.1 and 176.3 µg/mL, respectively, as determined by the MTT assay, while exerting negligible toxicity toward normal Vero cells. Miswak extract enhanced intracellular ROS production, disruption of MMP, nuclear condensation, and increased apoptotic cell populations, along with S-phase cell cycle arrest, pointing toward activation of mitochondrial-mediated apoptosis. In silico docking results indicated that key phytoconstituents exhibit strong binding interactions with multiple breast cancer-relevant targets such as ERα, PR, EGFR, HER3, IGF-1R, and GPER. Additionally, pharmacokinetic and toxicity predictions suggested favorable drug-like properties with minimal safety concerns. Thus, these findings support its potential as a promising plant-derived therapeutic candidate for breast cancer.