In vitro cytotoxicity and phytochemical analysis of the crude extract from Hypoxis hemerocallidea and its apoptosis-inducing effect on breast and skin can.

Hypoxis hemerocallidea, a medicinal plant traditionally used for its therapeutic properties, has demonstrated potential anticancer activity in vitro. This study evaluated the cytotoxic and pro-apoptotic effects of methanol (MeOH) and aqueous extracts of H. hemerocallidea on A375 melanoma, MCF-7 breast cancer, and HEK293 normal cell lines. MTT assays revealed concentration-dependent inhibition of cancer cell proliferation, with IC50 values of 44.82-63.1 µg/mL for MeOH extract and 55.02-57.6 µg/mL for aqueous extract, while sparing normal HEK293 cells. Morphological analysis showed characteristic apoptotic changes, including cell shrinkage, rounding, membrane blebbing, and formation of apoptotic bodies. Flow cytometric analysis demonstrated increased caspase-3 activity (13-15% for extracts vs. 47.8% for Doxorubicin) and mitochondrial membrane depolarization (17.4% for aqueous extract, 48.4% for MeOH extract vs. 56.5% for Doxorubicin), indicating activation of intrinsic apoptotic pathways. Collectively, these findings suggest that H. hemerocallidea extracts selectively induce apoptosis in cancer cells through mitochondrial-dependent mechanisms, highlighting their potential as natural anticancer agents. Further studies are warranted to isolate active compounds and elucidate their molecular targets.