Phytochemical Modulation of Fibrosis-Linked Oncogenic Pathways in Hepatocellular Carcinoma Cells

Abstract

Background:Liver fibrosis is a progressive pathological process that is the super-accumulation of extracellular matrix (ECM) due to chronic liver damage. Dysregulated signaling pathways involving interleukin-6 (IL-6), epidermal growth factor receptor (EGFR), and protein kinase B (AKT1), are implicated in hepatic stellate cell (HSC) activation, fibrogenesis, and progression toward cirrhosis and hepatocellular carcinoma (HCC). Despite advances in understanding these mechanisms, effective pharmacological interventions with favorable safety profiles remain limited. Methods:The current research examined the effects of two naturally occurring flavonoids, Kaempferol and Coumestrol on fibrosis-linked oncogenic signaling using an integrated in silico and in vitro study. Molecular docking studies were conducted to assess the binding affinities and the interaction profiles of both compounds with IL-6, EGFR, AKT1, and Caspase-3 (CASP3). Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) profiling was used to predict pharmacokinetic and toxicity properties to evaluate drug-likeness and safety parameters. In vitro experiments were conducted in HepG2 and Huh7 hepatocellular carcinoma cell lines to evaluate cytotoxicity, cell viability, migration, invasion, colony formation, and expression of IL-6, EGFR, AKT1, and Caspase-3. IC50 values were derived from dose-response curves generated from three independent technical experimental replicates.Results:Docking analysis showed stable binding interactions of both compounds at the active sites of IL-6, EGFR, AKT1, and CASP3, supported by hydrogen bonding and hydrophobic interactions, and ADMET predictions indicated favorable absorption with manageable toxicity profiles. In HepG2 and Huh7 cells, Kaempferol and Coumestrol showed dose-dependent reductions in cell viability, migration, invasion, and colony formation, along with downregulation of EGFR, AKT1, and IL-6 and upregulation of Caspase-3 at the transcript level. Conclusion: These findings suggest that Kaempferol and Coumestrol modulate IL-6/EGFR/AKT1-associated oncogenic signaling in hepatocellular carcinoma cells in vitro, indicating possible relevance to fibrosis-to-HCC progression. However, this study did not directly assess hepatic stellate cell activation or classical fibrogenic markers, and further studies using HSC models and in vivo validation are needed to confirm anti-fibrotic potential.