Brusatol induces ferroptosis to inhibit hepatocellular carcinoma progression by targeting ATF3

Ferroptosis is a novel form of programmed cell death that is triggered by iron‐dependent lipid peroxidation. Brusatol (BRU), a natural nuclear factor erythroid 2‐related factor 2 inhibitor, exhibits potent anticancer effects in various types of cancer. However, the exact mechanism of BRU in the treatment of hepatocellular carcinoma (HCC) remains unknown. The anticancer effects of BRU in HCC were detected using cell counting kit‐8 and colony formation assays and a xenograft model. RNA sequencing (RNA‐seq) and bioinformatics analyses of HCC cells were utilized to elucidate the mechanism underlying the effects of BRU in HCC. The levels of reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and Fe2+ were measured using assay kits. The expression of activating transcription factor 3 (ATF3) was tested using RT‐qPCR, western blotting, and immunofluorescence staining. The role of ATF3 in BRU‐induced ferroptosis was examined using siATF3. BRU significantly inhibited HCC cell proliferation, both in vitro and in vivo. BRU activated the ferroptosis signaling pathway and increased ATF3 expression. Furthermore, ATF3 knockdown impeded BRU‐induced ferroptosis. BRU suppressed HCC growth through ATF3‐mediated ferroptosis, supporting BRU as a promising therapeutic agent for HCC. In this study, we explored the mechanism of action of brusatol in hepatocellular carcinoma. Using RNA‐seq and through in vitro and in vivo studies, we determined that brusatol suppresses hepatocellular carcinoma progression by inducing ATF3‐mediated ferroptosis. Therefore, our research revealed the biological effect of brusatol treatment and provided ATF3 as a novel therapeutic target and prognostic biomarker for HCC therapy.