Cryptotanshinone Targets Ferroptosis in Glioma via the EGFR/ROS Signaling Pathway

To investigate the regulatory effects of cryptotanshinone (CTS) on the biological behavior of glioma cells and its underlying molecular mechanisms, with a particular focus on the role of the epidermal growth factor receptor/reactive oxygen species (EGFR/ROS) pathway in ferroptosis-mediated antitumor activity. Glioma cell behaviors were monitored through Cell Counting Kit-8 (CCK-8), colony formation, wound healing, and Transwell assays. Intracellular and tumor tissue levels of reactive oxygen species (ROS), mitochondrial membrane potential (MMP), ferrous iron (Fe²⁺), glutathione/glutathione disulfide (GSH/GSSG) ratio, and malondialdehyde (MDA) were assessed. Glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), acyl-CoA synthetase long-chain family member 4 (ACSL4), epidermal growth factor receptor (EGFR), and Ki67 expression was examined using western blotting and immunohistochemistry. Network pharmacology and molecular docking were employed to predict potential cryptotanshinone targets. An in vivo glioma model was created by implanting tumor cells into nude mice. CTS inhibited glioma cell malignant phenotype, while promoting ROS accumulation, MMP loss, Fe²⁺ elevation, and GSH depletion. CTS also modulated ferroptosis-associated molecules, characterized by downregulation of GPX4 and SLC7A11 and upregulation of ACSL4. EGFR was identified as a central target, which was experimentally validated to mediate the antitumor and ferroptosis-inducing effects of CTS. In vivo, CTS suppressed tumor growth and activated ferroptosis, whereas EGFR overexpression partially reversed these protective effects. CTS induces ferroptosis in glioma cells by inhibiting EGFR and enhancing ROS signaling, thereby suppressing tumor proliferation and invasion.