Assessment of Ferroptotic Anti-Cancer Potential of Ammonium Ferric Citrate and its Associated Biochemical Changes in Mammary Cancer MDA-MB-231 Cell Line

Cancer treatments have long focused on inducing apoptosis, but many cancer cells develop resistance by altering apoptotic protein levels, reducing drug effectiveness and causing side effects in healthy cells. These challenges have led researchers to explore alternative cell death mechanisms, particularly ferroptosis. This iron-dependent process, characterized by lipid peroxide accumulation, offers a promising approach for cancer therapy. In the present study, we evaluated the anticancer potential of a food additive ammonium ferric citrate (AFC) in MDA-MB-231 cell line. Cytotoxicity effect of AFC was evaluated by 3-(4,5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay and IC₅₀ value was found to be 0.01 mg/mL. Upon treatment with 0.01 mg/mL AFC intracellular iron levels were estimated using the Cell Ferrous Iron Colorimetric Assay Kit, Reduced glutathione (GSH) concentration, Superoxide dismutase (SOD) activity, total antioxidant capacity (TAC) malondialdehyde (MDA) and reactive oxygen species (ROS) in MDA-MB-231 cells were quantified as per standard protocol. AFC treatment significantly increased intracellular iron MDA and ROS levels compared to untreated cells (P < 0.05). While SOD activity, GSH level and TAC significantly (P < 0.05) decreases upon treatment with AFC. Our findings highlight the potential of AFC as an inducer of ferroptosis in TNBC cells by promoting iron overload, oxidative stress, and lipid peroxidation. These results suggest that AFC could be explored as a candidate for ferroptosis-based cancer therapy, warranting further investigation in preclinical and in vivo models.