ABSTRACT: Background & Aims: Hepatocellular carcinoma (HCC) is the most prevalent type of primary liver cancer.Globally, liver cancers represent the fourth most common cause of cancer-related deaths. Although statins have been shown to be effective in preventing HCC, their underlying molecular mechanisms are not yet fully elucidated. The aim of this study was to evaluate the hepatic gene expression profile and its relationship to early hepatocarcinogenic changes in a Hexachlorobenzene (HCB)-induced preneoplastic liver lesion model in rats treated with atorvastatin (AT). Materials & Methods: Male Wistar rats weighing 160 g b.w. were randomly assigned to four experimental groups ( n= 5 per group) : control, HCB (100 mg/kg b.w.), AT (5 mg/kg b.w.), and AT + HCB, treated for 30 days. The analysed parameters were liver weight, body weight, hepatosomatic index, liver histology, and expression levels of target proteins, c-Myc, TGF-β1, and p53 by immunofluorescence (n = 3 per group). Transcriptomic analysis was performed by RNA sequencing on liver tissue (n = 2 per group). Differentially expressed genes (DEGs) were identified using DESeq2 (|log2FC| > 1, adjusted p < 0.05).. Gene set enrichment analysis (GSEA) and KEGG pathway overrepresentation analysis were conducted using the WebGestalt platform . Results: HCB treatment significantly increased the hepatosomatic index (28%, p<0.01) and induced preneoplastic hepatic lesions, both effects were prevented by AT co-administration. Transcriptomic analysis of HCB-treated livers identified 6,396 DEGs, of which 1,352 overlapped with the 1,612-gene KEGG “Pathway in cancer” set (83.9% concordance; p = 5.19×10⁻⁵⁴, FDR = 1.82×10⁻⁵¹). HCB activated multiple oncogenic pathways, including RHO GTPase cycle, TGF-β signalling, and receptor tyrosine kinase signalling, AT co-treatment downregulated these HCB-induced pathways and upregulated protective PPAR signalling, autophagy, and cellular stress response pathways. Consistent with these transcriptomic findings, AT co-administration reduced TGF-β1 and c-Myc protein levels and attenuated the HCB-induced accumulation of total p53 protein levels observed in liver tissue. Conclusions: In this experimental model, Atorvastatin exerted a protective effect against HCB-induced preneoplastic hepatic changes through the modulation of oncogenic and cytoprotective transcriptomic programmes. These findings provide mechanistic insights into statin-mediated hepatoprotection and support further investigation of AT as a potential chemopreventive agent in high-risk populations. However, additional validation in models that better reflect the chronic liver injury context of human HCC is needed.
