Apigenin Inhibits Triple‐Negative Breast Cancer Growth by Dual Targeting GPX4 / SLC7A11 ‐Mediated Ferroptosis and PKM2 / GLUT1 ‐Mediated Aerobic Glycolysis

Triple-negative breast cancer (TNBC) is aggressive with limited treatments. Although the natural flavonoid Apigenin (API) shows anti-tumor potential, its mechanism in TNBC remains unclear. This study investigated API's role in inducing ferroptosis and inhibiting glycolysis to suppress TNBC. Molecular docking predicted API's binding to ferroptosis- and glycolysis-related proteins. In vitro, 4T1 and MDA-MB-231 TNBC cells were used to assess API's effects on viability, migration, invasion, and key metabolic markers (Fe2+, MDA, ROS, GSH, lactic acid, glucose, ATP, OCR, ECAR), and protein expression (GPX4, SLC7A11, TFR, FPN1, FTH1, FTL, FSP1, PKM2, GLUT1, GLUT4, HK2, LDHA) using RT-qPCR and Western blotting. In vivo, a 4T1 tumor xenograft model evaluated API's impact on tumor growth, protein expression, and toxicity (H&E staining). Molecular docking indicated good binding affinity of API with ferroptosis- and glycolysis-related proteins. In vitro, API inhibited the viability, migration, and invasion of TNBC cells. API significantly increased Fe2+, MDA, and ROS levels while decreasing reduced GSH levels, downregulated GPX4, SLC7A11, FPN1, FTH1, FTL, and FSP1 expression, and upregulated TFR expression, indicating ferroptosis induction. API also decreased lactic acid, ATP, and ECAR levels while increasing intracellular glucose and OCR levels, downregulating PKM2, GLUT1, GLUT4, HK2, and LDHA expression, demonstrating glycolysis inhibition. In vivo, API significantly inhibited tumor growth in the 4T1 xenograft model without obvious toxicity and regulated the expression of ferroptosis and glycolysis-related proteins. These findings establish API as a promising natural anti-TNBC drug by simultaneously targeting ferroptosis and aerobic glycolysis.