Abstract 1614: Ferroptosis mediated cell death in colorectal cancer cells through oxidative stress

Abstract Ferroptosis, a regulated form of non-apoptotic cell death, occurs due to lipid peroxide accumulation resulting from impaired cellular antioxidant defenses, particularly the inactivation of glutathione peroxidase 4 (GPX4). This process has gained attention as a novel therapeutic target, particularly for colorectal cancer (CRC), a malignancy often resistant to conventional apoptosis-based therapies. Our study investigated the anticancer potential of HLN, an antimicrobial agent. The cytotoxicity of HLN was evaluated in CT26, HCT116 & HT29 cell lines by SRB assay. To confirm HLN-induced ferroptosis, we employed ferro-orange staining to detect lipid peroxidation, GSH and malondialdehyde (MDA) assays to quantify oxidative damage. Mitochondrial membrane potential and ROS generation were determined by flow cytometry, Western blotting was used to determine GPX4, GPX1, NRF2, KEAP1 and SLC7A11/xCT expression in CRC cells. HLN exhibited significant cytotoxic effects across all CRC cell lines, with IC50 values ranging from 1-4 μM. Ferro-orange staining and MDA assays confirmed extensive lipid peroxidation by HLN treatment. HLN treatment also significantly depleted intracellular GSH levels and depolarized mitochondrial membrane, whereas increased ROS production, indicating oxidative stress. Western blot results showed reduced GPX4 and SLC7A11/xCT as well as modulation in ferroptotic markers, such as GPX1, NRF2, KEAP1, by HLN treatment. In conclusion our data suggests that HLN induces ferroptosis in CRC cells by promoting lipid peroxidation, mitochondrial dysfunction, and oxidative stress Citation Format: Mohamed A. Eltokhy, Shreyas R. Gaikwad, Morgan Moore, Sanjay K. Srivastava. Ferroptosis mediated cell death in colorectal cancer cells through oxidative stress [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1614.