Evaluation of lipid peroxidation as a marker of ferroptosis in colon cancer cells.

Ferroptosis is a regulated form of cell death driven by iron-dependent lipid peroxidation, with emerging relevance in cancer biology, particularly in therapy-resistant tumors such as colorectal cancer. Accurate and reproducible methods to monitor ferroptosis are essential for understanding its mechanisms and therapeutic potential. In this methodological paper, we present an optimized protocol for the evaluation of lipid peroxidation as a surrogate marker of ferroptosis in colon cancer cell models. Specifically, we describe an approach for quantifying malonylaldehyde and hydroxyalkenals (MDA+ 4-HDA), as key byproducts of lipid oxidation, using a colorimetric assay adapted for cell culture. This methodological framework provides a reliable basis for dissecting ferroptotic responses and for evaluating the activity of ferroptosis-inducing compounds in colorectal cancer research.