# Conjugated Linolenic Acids Induce Ferroptosis in Human and Zebrafish Melanoma Cells

## TL;DR

Conjugated linolenic acids trigger ferroptosis in melanoma cells from humans and zebrafish, offering a new approach to treat this aggressive cancer.

## Contribution

This study identifies specific conjugated linolenic acid isomers as potent inducers of ferroptosis in melanoma cells.

## Key findings

- Conjugated linolenic acids reduced cell viability in human and zebrafish melanoma cells in a dose-dependent manner.
- Ferroptosis inhibitors mitigated cell death, confirming ferroptosis as the primary mechanism.
- CLnAs were incorporated into phospholipids, promoting lipid peroxidation and upregulating pro-ferroptotic genes.

## Abstract

Conjugated linolenic acids (CLnAs) are emerging as promising agents to trigger ferroptosis, a cell death driven by excessive lipid peroxidation, in cancer cells. Given the aggressive nature and treatment resistance of malignant melanoma, exploring CLnAs as therapeutic agents may offer a novel strategy to overcome these challenges. Here, we investigated the toxicity of four CLnA isomers on human (A375, WM266.4) and zebrafish (ZMEL1) melanoma cell lines. We observed a dose-dependent reduction in cell viability across all three tested cell lines. While human melanoma cells were more sensitive to CLnAs than ZMEL1 cells, treatment with ferroptosis inhibitors mitigated cell death in all models, confirming ferroptosis as the consistent primary mechanism of cell death. In addition, chemical inhibitors of ACSL4 and GPX4 modulated CLnA toxicity, further substantiating the ferroptotic mechanism by highlighting the role of these key regulators. Furthermore, fatty acid analysis revealed that CLnAs were effectively incorporated into phospholipids, generating substrates for lethal lipid peroxidation. At the transcriptional level, CLnA treatment significantly upregulated the pro-ferroptotic gene acsl4a in ZMEL1 cells. Overall, our study identifies specific CLnAs as potent ferroptosis inducers in both human and zebrafish melanoma cells and underscores the translational relevance of the zebrafish model based on a shared ferroptotic mechanism.