Transcriptomic Analysis Reveals Core Genes and Mechanisms Linking Grilled Food Toxicants (4-HNE, BaP, PhIP) to NASH-HCC Progression.

Grilled food toxicants 4-hydroxynonenal (4-HNE), benzo[a]pyrene (BaP), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) pose health risks, including non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). However, their synergistic molecular mechanisms in driving NASH to NASH-HCC progression remain unclear. We employed network toxicology (CTD, GeneCards), bioinformatics analysis of the GSE164760 dataset, machine learning (LASSO, SVM-RFE, Random Forest), SHAP interpretability analysis, immune infiltration profiling (CIBERSORT), miRNA-TF-mRNA network construction, and molecular docking to identify core genes and elucidate mechanisms. Toxicity prediction confirmed significant hepatotoxicity for all three compounds. Intersection analysis identified 8 key genes (InterGenes) enriched in oxidant detoxification, fatty acid metabolism, and chemical carcinogenesis pathways. Machine learning refined this to 6 CoreGenes (GSTA1, EPHX1, CYP2E1, GPX3, ALB, TF). SHAP analysis revealed that high expression of ALB, TF, GSTA1 and CYP2E1, together with low expression of GPX3 and EPHX1, correlated with increased progression risk. A CoreGenes-based diagnostic nomogram achieved exceptional performance (AUC = 0.967). Immune infiltration analysis revealed significant shifts in regulatory T cells and M2 macrophages during progression. Molecular docking confirmed strong binding affinities between the toxicants and CoreGenes proteins (e.g., BaP-GSTA1: -10.7 kcal/mol). Functional enrichment implicated dysregulated fatty acid metabolism, PPAR, and TGF-beta signaling. This study identifies GSTA1, EPHX1, CYP2E1, GPX3, ALB, and TF as pivotal biomarkers and mediators through which 4-HNE, BaP, and PhIP synergistically promote NASH-HCC progression via oxidative stress, metabolic dysfunction and remodeling of the immune microenvironment. The CoreGenes signature and multi-gene model provide a powerful tool for risk assessment and early intervention.