In vitro anticancer efficacy of Asparagus khorasanensis Extract on MDA-MB-231 Cells for Enhanced Apoptosis Induction in Breast Cancer

Natural plant-derived compounds are emerging as promising alternatives to conventional chemotherapy due to their selective toxicity and reduced side effects. Asparagus khorasanensis , a traditionally used medicinal plant, remains unexplored for its anticancer potential. This study investigated the cytotoxic and pro-apoptotic effects of a hydroalcoholic extract of A. khorasanensis on triple-negative MDA-MB-231 breast cancer cells, alongside its safety profile in normal human embryonic kidney cells (HEK-293). The extract was chemically characterized by gas chromatography–mass spectrometry (GC-MS), and its cytotoxicity was assessed via MTT assay across various concentrations (3, 6.25, 12.5, 25 and 50 μg/ml) and time points (24 h, 48 h and 72 h). Apoptosis was evaluated by Annexin V-FITC/PI flow cytometry, and expression of apoptosis-related genes ( CASPASE-3 , CASPASE-8 , BAX , BAK , P53 , BCL-2 ) was analyzed by real-time PCR. GC-MS identified Carvacrol and p-thymol as major constituents, reported for the first time in this species. The extract induced significant, concentration- and time-dependent cytotoxicity in MDA-MB-231 cells (IC₅₀ = 24.7 µg/ml at 48 h, p < 0.05), while exhibiting negligible effects on HEK-293 cells. Flow cytometry confirmed increased apoptotic populations, and gene expression analysis revealed upregulation of pro-apoptotic markers and downregulation of BCL-2 . These findings provide new evidence for the selective anticancer activity of A. khorasanensis extract and support further investigation of its active constituents as potential complementary therapeutic agents. • Asparagus khorasanensis extract induces apoptosis in MDA-MB-231 breast cancer cells. • The extract shows minimal cytotoxicity in normal HEK-293 cells. • Apoptosis was confirmed by flow cytometry and qPCR ( BAX, BAK, P53, BCL-2 , CASPASE-3 , CASPASE-8 ).