TRIP13 Promotes Aflatoxin B1-Induced Hepatocellular Carcinoma Progression by Inhibiting Ferroptosis

Abstract

Aflatoxin B1 (AFB1) is a potent foodborne hepatocarcinogen that contributes substantially to the global burden of hepatocellular carcinoma (HCC), yet the molecular mechanisms underlying AFB1-driven HCC progression remain incompletely understood. In this study, we identified thyroid hormone receptor-interacting protein 13 (TRIP13) as an important contributor to AFB1-related HCC progression and investigated its involvement in ferroptosis regulation. TCGA-LIHC, GSE25844, and single-cell RNA sequencing data (GSE166635) revealed that TRIP13 was significantly upregulated in AFB1-related HCC, enriched in malignant cell populations, and associated with poor clinical outcomes. In Huh7 cells, AFB1 treatment markedly increased TRIP13 expression, while TRIP13 knockdown significantly suppressed AFB1-induced cell proliferation, colony formation, migration, and epithelial-mesenchymal transition (EMT). Bulk RNA sequencing and pathway enrichment analyses indicated that TRIP13 overexpression was strongly associated with ferroptosis-related signaling. Mechanistically, TRIP13 was associated with ferroptosis resistance through modulation of the KEAP1/NRF2/xCT/GPX4 pathway, thereby contributing to redox homeostasis during AFB1 exposure. Consistently, TRIP13 knockdown restored ferroptotic sensitivity, as evidenced by increased Fe²⁺ and malondialdehyde levels and altered glutathione metabolism. Importantly, ferroptosis induction with RSL3 effectively reversed the pro-tumor effects of TRIP13 overexpression both in vitro and in vivo, including enhanced proliferation, EMT, migration, and xenograft tumor growth. Collectively, these findings suggest that TRIP13 contributes to AFB1-associated HCC progression through ferroptosis-related regulation and highlight the AFB1/TRIP13/ferroptosis axis as a potential molecular mechanism and candidate target for further investigation in AFB1-related HCC.