Abstract
Aristolochic acid I (AAI) is a known liver carcinogen, but its molecular mechanisms in hepatocellular carcinoma (HCC) remain incompletely understood. This study aimed to systematically elucidate these mechanisms. This study employed a multidisciplinary strategy: assessing AAI carcinogenicity via ProTox-3.0; identifying its targets using PubChem; and screening HCC-related genes from GEO. Core genes were filtered through network toxicology and Lasso-Cox regression. Molecular docking and dynamics simulations analyzed AAI-protein interactions, while GO, KEGG, and GSEA enrichment revealed relevant pathways. Key gene expression and clinical relevance were validated via TCGA and HPA databases. The carcinogenic potential of the AAI was 0.77. Fifty-two potential targets of AAI-induced HCC were identified. Network toxicology analysis identified 12 core genes including EZH2, FABP5, and RXRA. Lasso-Cox regression analysis identified four core genes that drive HCC progression, including EZH2. Functional enrichment analysis revealed that AAI-HCC targets were significantly enriched in lipid metabolism and the PPAR signaling pathway (p < 0.05), with the PPAR signaling pathway being significantly activated in HCC. Molecular docking and MD simulations confirmed the high affinity and stable binding of AAI to EZH2, FABP5, and RXRA. Through in silico and database-based validation, biological samples and clinical data confirmed that these genes were significantly overexpressed in HCC (p < 0.001) and were correlated with poor prognosis (p < 0.01). AAI may synergistically activate the PPAR signaling pathway by targeting EZH2, FABP5, and RXRA, driving lipid metabolism reprogramming and promoting the occurrence and development of HCC.