Characterizing Molecular Pathways of Precancerous Changes in Hepatocytes Under Metabolic Overload

Background Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths, often diagnosed late with a poor prognosis. This study aimed to elucidate molecular processes associated with precancerous alterations arising from metabolic overload in hepatocyte like cells, shedding light on potential pathomechanisms underlying non-cirrhotic NAFLD progression and hepatocarcinogenesis.

Methods Differentiated HepaRG cells, as modell of unaltered hepatocytes, were subjected to in vitro treatments mimicking metabolic syndrome by exposing them to excess insulin, glucose, and free fatty acids (FFA) up to 48h. mRNA expression of key genes involved in fat and glucose metabolism, and the tumor related genes PTEN, TP53, Abl1, ANXA2-4, and SIRT1were quantified via real-time PCR. Intracellular protein expression and phosphorylation were analyzed through Western blotting for genes related to cell cycle control (PCNA, E2F) and PI3K signaling (NF-kappaB, PTEN).

Results Robust cellular steatosis of fully differentiated HepaRG cells was detected under treatment with FFA and combined glucose, insulin, and FFA treatment. However, no significant changes in mRNA expression of the analyzed genes was detected.

Conclusion Short term steatosis seems not to affect expression or regulation of genes involved in fat or glucose metabolism, tumor suppression, or growth regulation. Long-term effects of metabolic overload should be analyzed in appropriate models to estimate its impact on non-cirrhotic tumorigenesis.

Publication History

Article published online:
23 January 2024