Genistein Induces Ferroptosis in Colorectal Cancer Cells via FoxO3/SLC7A11/GPX4 Signaling Pathway

Colorectal cancer is among the most frequently diagnosed cancers with high mortality rates and poses a serious threat to human health. Genistein (Gen) has been found to have anti-colorectal cancer effects, however, the molecular mechanisms by which genistein elicits its effects on colorectal cancer (CRC) cells have not been fully elucidated. In this study, we investigated the oxidative state of colorectal cancer cells during the antitumor action of Genistein and whether it can exert its antitumor effects through ferroptosis. Current research on the oxidative state of Genistein indicates that it exhibits both antioxidant and pro-oxidant properties. Different drug concentrations were applied to colorectal cancer cells, after which cell viability and key markers of ferroptosis, including reactive oxygen species (ROS), malondialdehyde (MDA), and Fe²⁺, were measured. We found that genistein significantly reduced the viability of colorectal cancer cells, and the expression of ferroptosis markers increased in a concentration-dependent manner. Subsequently, we treated cells with the ferroptosis inhibitor fer-1 in combination with genistein and observed a partial reversal of ferroptosis markers. These findings suggest that genistein exerts its antitumor effect by promoting iron-dependent oxidative damage-induced ferroptosis. To further elucidate the mechanism underlying ferroptosis modulation, we examined the protein and mRNA expression levels of the classical key ferroptosis molecules SLC7A11 and GPX4. We found that the expression levels of these molecules decreased, with GPX4 exhibiting a greater decrease. Overexpression of GPX4 reversed the pro-ferroptotic effect of genistein, indicating that genistein promotes ferroptosis occurrence by downregulating GPX4 expression. When the drug was applied to colorectal cancer cells, the expression of the transcription factor FoxO3 increased. Treatment of cells with the FoxO3 inhibitor JY-2 in combination with other drugs resulted in antagonism of ferroptosis markers. These findings suggest that genistein induces ferroptosis in colorectal cancer cells through the FoxO3/SLC7A11/GPX4 signaling pathway, thereby inhibiting tumor growth.