ONC212 Induces Ferroptosis-Associated Cell Death in Glioblastoma Cells

Abstract

Aim: Glioblastoma (GBM) is an aggressive primary brain tumor with limited treatment options. Ferroptosis, an iron-dependent cell death characterized by lipid peroxidation and oxidative stress, is a promising therapeutic target. This study investigated the anticancer effects of the imipridone derivative ONC212 in GBM cells and examined whether its cytotoxicity involves ferroptosis-related mechanisms.Material and Methods: U87 and U251 cells were treated with increasing ONC212 concentrations. Viability was assessed by CCK-8, proliferation by BrdU incorporation (24–72 h). Ferroptosis-associated changes were evaluated by measuring ferrous iron (Fe²⁺), malondialdehyde (MDA), glutathione (GSH), and reactive oxygen species (ROS). Expression of ferroptosis-related genes GPX4 and ACSL4 was analyzed by qRT-PCR. Ferrostatin-1 (Fer-1) was used to confirm ferroptosis involvement.Results: ONC212 reduced viability in a time- and dose-dependent manner [IC₅₀ (95% CI): 18.7 µM (17.3-20.1), 9.6 µM (8.8-10.4), and 6.2 µM (5.7-6.7) at 24, 48, and 72 h, respectively]. BrdU incorporation decreased by 46.9% and 79.4% at 24 h and 72 h, respectively, following treatment with the 24-hour IC₅₀ concentration (18.7 µM). Ferroptosis-associated oxidative stress was evident through increased Fe²⁺, MDA, ROS, and GSH depletion. ONC212 upregulated ACSL4 (2.14‑fold) and reduced GPX4 expression by 62%. Fer-1 pretreatment partially reversed these alterations, supporting the involvement of ferroptosis-associated mechanisms. Similar results were obtained in U251 cells.Conclusion: ONC212 exerts potent cytotoxic and anti-proliferative effects in GBM cells and provides preliminary evidence suggesting that it induces ferroptosis-associated cell death through modulation of oxidative stress and ferroptosis‑associated gene expression. Targeting ferroptosis with ONC212 may represent a promising therapeutic strategy for GBM.