Hepatocellular carcinoma (HCC) is one of the most prevalent and lethal
cancers worldwide and lacks effective treatment. Herein, we found that the antifungal
Natamycin (
NAT) exhibits antitumor activity by inducing apoptosis both in vitro and in vivo. Mechanistically,
NAT downregulates the expression of
Peroxiredoxin 1 (PRDX1) by promoting ubiquitination-mediated degradation, thereby leading to increased
reactive oxygen species (ROS) accumulation and subsequent apoptosis. Exogenous overexpression of PRDX1 or
N-acetyl-l-cysteine (NAC) pretreatment abrogates
NAT-induced cytotoxicity in PLC/PRF/5 and Huh7 cells, suggesting the vital role of ROS in the antitumor properties of
NAT. Of note, downregulation of PRDX1 decreases the phosphorylation of AKT, thereby inducing cytoprotective autophagy and combinational use of
NAT and
chloroquine (CQ) achieves better anti-
tumor efficacy. Moreover,
NAT acts synergistically with
sorafenib (SOR) in HCC suppression. Collectively, our study provides an important molecular basis for
NAT-induced cell death and suggests that the antifungal
NAT holds the potential to be repurposed as an anticancer
drug for HCC treatment.