Ex vivo evaluation of Allium sativum extract on acute myeloid leukemia cells and leukemia stem cell populations.

Despite therapeutic advances, acute myeloid leukemia (AML) remains associated with high relapse rates and poor outcomes, especially in elderly or unfit patients. Allium sativum is widely known for its medicinal properties, but its anti-leukemic effects have not been fully explored. This study investigated the cytotoxic and pro-apoptotic potential of an ethyl acetate extract of Allium sativum (EAEAS) using ex vivo mononuclear cells from thirteen AML patients. Cytotoxicity was assessed via MTT assay, while mechanisms of cell death were analyzed using annexin V/propidium iodide staining, DNA fragmentation, mitochondrial depolarization, and caspase 3/7 activation. EAEAS induced a dose-dependent decrease in cell viability, with variable IC₅₀ values across samples. Apoptosis was confirmed through mitochondrial dysfunction, sub-G1 accumulation, and caspase activation. Additionally, we evaluated the effect of EAEAS on two LSC subpopulations using multicolor flow cytometry. Although no statistically significant changes were observed, some patients showed a decrease in LSC frequencies after treatment, suggesting possible selective activity. These findings indicate that EAEAS triggers intrinsic apoptosis in AML blasts and may exert partial activity on stem-like compartments, warranting further evaluation as an adjuvant therapy in AML.