Short‐Chain Fatty Acids Modulate Iron Metabolism and Induce Oxidative Stress and Ferroptosis in Breast Cancer Cells

Short-chain fatty acids (SCFAs), produced through the fermentation of dietary fibers by gut microbiota, play a crucial role in regulating gut microbiota composition, immune function, and overall health. Emerging evidence suggests that SCFAs exhibit antiproliferative effects in various cancers, though the underlying molecular mechanisms remain unclear, particularly in breast cancer (BC). This study aimed to explore the impact of SCFAs, including sodium acetate, propionate, butyrate, and pentanoate, on iron metabolism and ferroptosis in BC cells. The effects of SCFAs were examined in MCF-7 and MDA-MB-231 BC cell lines. Cell viability, intracellular iron levels, and expression of iron regulatory proteins transferrin (TfR1), ferroportin (FPN), hepcidin, and ferritin (FTH1) were analyzed. Additionally, oxidative stress markers such as reactive oxygen species (ROS), lipid peroxidation, glutathione peroxidase 4 (GPX4), and mitochondrial membrane potential were assessed. Adenosine triphosphate (ATP) production was also measured to determine ferroptosis involvement. SCFAs exhibited a dose-dependent reduction in BC cell proliferation. They increased intracellular iron content, altered iron metabolism-related protein expression, and elevated oxidative stress. This was evidenced by increased ROS levels, lipid peroxidation, GPX4 downregulation, mitochondrial membrane depolarization, and ATP depletion. SCFAs modulate iron metabolism in BC cells, triggering oxidative stress and ferroptosis-induced cell death. These findings highlight SCFAs' potential as therapeutic agents in BC treatment.