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Analysis of gene expression changes in lipid droplet formation of human liver cancer cell HepG2

ABSTRACT: Fatty liver is a condition of excessive triglyceride accumulation in hepatocytes. Additionally, hepatocytes showed a high degree of fat droplet accumulation during excessive alcohol consumption and metabolic syndrome. However, the molecular mechanisms involved in fat droplet formation remain unknown. This study used an in vitro fatty liver formation model of the human liver cancer cell line HepG2 to comprehensively search for fat droplet formation-related genes whose expression changes during fat droplet formation. Microarray analysis with extracted total RNA determined the genes that are involved in fat droplet formation and confirmed their expression by real-time polymerase chain reaction after culturing HepG2 cells in a culture medium containing 0, 50, 200, and 500 μM of oleic acid for 24 h. The results revealed 142 genes demonstrating increased expressions by >2.0 fold with oleic acid treatment and 426 genes demonstrating decreased expressions. Perilipin 2 (PLIN2) was estimated as the gene most closely associated with fatty liver. Lipid droplet formation in HepG2 upregulated PLIN2 in an oleic acid-treated concentration-dependent manner. These results indicate the involvement of genes in oleate treatment-induced lipid droplet formation in HepG2, particularly PLIN2 may play an important role.

ORGANISM(S): Homo sapiens

PROVIDER: GSE248166 | GEO | 2023/12/01

REPOSITORIES: GEO

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Project description:How hepatocytes respond to a fatty acid stimulus is the topic of continuous research, since it is essential for our understanding of non-alcoholic fatty liver disease a pathology with a hard to detect onset, estimated to be present in a quarter of the adult human population. As advanced stages include cirrhosis and development of hepatocellular carcinoma, early detection and intervention is crucial. To improve our understanding of the development of non-alcoholic fatty liver disease we treated a human hepatoma cell line model, HepG2, with increasing concentrations of common fatty acids namely myristic, palmitic and oleic acid. To reproduce more representative conditions, we also included combinations of these fatty acids and monitored the cellular response with an in-depth proteomics approach and imaging techniques. The two saturated fatty acids presented a similar phenotype of a dose dependent decrease of proliferation rates and little lipid droplet formation. Interestingly, our data corroborated that this drop in proliferation rates was due to delayed cell cycle progression following myristic acid treatment, whereas palmitic acid led to cellular apoptosis. In contrast, oleic acid, as well as saturated fatty acid mixtures with oleic acid, led to a dose dependent increase of lipid droplet volume without the adverse impact on proliferation. Comparing the outcomes from harmful single fatty acid treatments and the well tolerated fatty acid mixes, we were able to differentiate between fatty acid specific cellular responses and likely common lipotoxic denominators. Two of them, A2M and SERPINA3, are modulators of the innate immune system and are likely involved in the inflammation occurring on systemic level. The remaining four include transcription factors like H2AFY, which recently has been proposed as a biomarker for hepatocellular carcinoma. Conclusively, our study not only manages to contextualise proteome alterations following fatty acid treatment, but also highlights differences between cellular effects of two common saturated fatty acids.

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Analysis of gene expression changes in lipid droplet formation of human liver cancer cell HepG2

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