Antioxidant and Anticancer Effects of Alchemilla vulgaris Extract on Medullary Thyroid Carcinoma Cells

Abstract This study examined the antioxidant and anticancer properties of the ethyl acetate extract of Alchemilla vulgaris L. (A. vulgaris) against medullary thyroid carcinoma (MTC-TT) cells. Antioxidant activity was measured through assays of total antioxidant capacity, phenolic content, and reducing power. Cytotoxicity, apoptosis, proliferation, and migration were evaluated using MTT, TUNEL, colony formation, and wound-healing assays. Gene expression of CDKN1A, BAX, BCL-2, MMP2, and MMP9 was analyzed by RT-PCR. The extract showed strong antioxidant potential with high phenolic levels and notable reducing activity. Treatment of MTC-TT cells led to reduced viability in a time- and dose-dependent manner, with increased apoptosis, BAX induction, BCL-2 repression, and elevation of CDKN1A expression over time. Colony formation was significantly decreased, and wound healing assays confirmed reduced migration, which was consistent with transient downregulation of MMP2 and MMP9. Selectivity was evident, as IC₅₀ values were substantially higher in WI-38 normal fibroblasts compared to tumor cells. These findings suggest that A. vulgaris ethyl acetate extract exerts anticancer activity by influencing apoptosis, cell growth, and migration. Its phenolic richness and robust antioxidant profile highlight the extract as a promising nutraceutical candidate for further in vivo research in aggressive and treatment-resistant cancers such as MTC.