Evaluation of the Antioxidant Potential and Selective Cytotoxicity of Fumaria agraria Alkaloids Against Human Breast Cancer Cells

Abstract

This study aimed to investigate the antioxidant capacity, hemolytic effect, and cytotoxic potential of the alkaloid fraction isolated from Fumaria agraria , with an emphasis on its possible use as a therapeutic agent against cancer cells. The antioxidant activity of the alkaloidal fractions was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH), reducing power, and phosphomolybdate assays. Hemolytic activity was assessed to determine potential erythrocyte toxicity. Cytotoxicity was tested on Michigan Cancer Foundation-7 (MCF-7), Michigan Cancer Foundation-231 (MDA-MB-231), and Michigan Cancer Foundation-10A (MCF-10A) breast cell lines. Cell cycle distribution and cyclin expression were analyzed by flow cytometry, and apoptosis was detected by dual staining with propidium iodide (PI) and Annexin V. The basic fraction (BF) showed strong antioxidant activity (half-maximal inhibitory concentration [IC 50 ] = 50.34 µg/mL in the DPPH assay) and low hemolytic toxicity (4.1% at 2 mg/mL), lower than boldine (5.8%). It exhibited notable cytotoxicity against MCF-7 cells and moderate effects on MDA-MB-231 cells, while normal MCF-10A cells were less sensitive. Flow cytometry revealed G 0 /G 1 arrest in MCF-7 cells and G 2 /M accumulation in MDA-MB-231 cells. In MCF-7 cells, cyclin D1, cyclin E, and MYC proto-oncogene, bHLH transcription factor (MYC) expression decreased, whereas cyclin-dependent kinase 1 (CDK1)/cyclin B1 activity increased in MDA-MB-231 cells. Apoptosis occurred in MCF-7 cells but not in MDA-MB-231 cells. The alkaloid fraction from F. agraria displayed potent antioxidant and selective cytotoxic effects of BF, particularly against MCF-7 breast cancer cells, with minimal toxicity toward normal cells. These findings highlight its potential as a promising source of bioactive compounds for therapeutic development.