Anticancer potential of Allium sativum against triple-negative breast cancer cells: Evidence from ROS-mediated cell cycle arrest and apoptosis

Breast cancer is a common disease that impacts the women population worldwide and therefore requires effective treatment strategies. Allium sativum L. (A. sativum) has been consumed in conventional remedies since antiquity owing to its extensive medicinal value. This study set out to analyse the primary phytocomponents in the active fraction of A. sativum and to evaluate its in vitro anticancer potential against triple-negative breast cancer (TNBC) MDA-MB-231 cells. Nonpolar to polar fractions of A. sativum extract were subjected to MTT assay, and the active fraction was selected for ROS estimation, apoptosis studies, mitochondrial membrane potential (MMP) assessment and cell cycle analysis. The active fraction was characterized phytochemically using the HPLC technique. The ethyl acetate fraction from A. sativum (EAFAS) was identified as the active fraction based on the MTT screening test. HPLC investigation confirmed two main organosulfur compounds, allicin and alliin in EAFAS. EAFAS treated MDA-MB-231 cells exhibited apoptotic-like morphology; however, no any substantial cytotoxicity toward the normal epithelial cell line Vero was observed. Fluorescence microscope and flow cytometry analysis confirmed both apoptotic and necrotic cell death in TNBC cells. Notably, EAFAS showed a low amount of MMP and a high level of ROS production. A cell-cycle study showed that exposure to the EAFAS enhanced the proportion of cells in the S and G2/M stages. These findings provide crucial new insights into the potential of EAFAS as a medicinal candidate for the treatment of TNBC.