Although human liver tumor cells have reduced metabolic functions as compared to
primary human hepatocytes (PHH) they are widely used for pre-screening tests of drug metabolism
and toxicity. The aim of the present study was to modify liver cancer cell lines in order to improve
their drug-metabolizing activities towards PHH. It is well-known that epigenetics is strongly modified
in tumor cells and that epigenetic regulators influence the expression and function of Cytochrome
P450 (CYP) enzymes through altering crucial transcription factors responsible for drug-metabolizing
enzymes. Therefore, we screened the epigenetic status of four different liver cancer cell lines (Huh7,
HLE, HepG2 and AKN-1) which were reported to have metabolizing drug activities. Our results
showed that HepG2 cells demonstrated the highest similarity compared to PHH. Thus, we modified
the epigenetic status of HepG2 cells towards ‘normal’ liver cells by 5-Azacytidine (5-AZA) and Vitamin
C exposure. Then, mRNA expression of Epithelial-mesenchymal transition (EMT) marker SNAIL
and CYP enzymes were measured by PCR and determinate specific drug metabolites, associated
with CYP enzymes by LC/MS. Our results demonstrated an epigenetic shift in HepG2 cells towards
PHH after exposure to 5-AZA and Vitamin C which resulted in a higher expression and activity of
specific drug metabolizing CYP enzymes. Finally, we observed that 5-AZA and Vitamin C led to
an increased expression of Hepatocyte nuclear factor 4α (HNF4α) and E-Cadherin and a significant
down regulation of Snail1 (SNAIL), the key transcriptional repressor of E-Cadherin. Our study
shows, that certain phase I genes and their enzyme activities are increased by epigenetic modification
in HepG2 cells with a concomitant reduction of EMT marker gene SNAIL. The enhancing of liver
specific functions in hepatoma cells using epigenetic modifiers opens new opportunities for the usage
of cell lines as a potential liver in vitro model for drug testing and development.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:88964 |
| Date | 11 January 2024 |
| Creators | Ruoß, Marc, Damm, Georg, Vosough, Massoud, Ehret, Lisa, Grom-Baumgarten, Carl, Petkov, Martin, Naddalin, Silvio, Ladurner, Ruth, Seehofer, Daniel, Nussler, Andreas, Sajadian, Sahar |
| Publisher | MDPI |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:article, info:eu-repo/semantics/article, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 347, 10.3390/ijms20020347 |
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