Epigenetic Modifications of the Liver Tumor Cell Line HepG2 Increase Their Drug Metabolic Capacity

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-metabolizin...

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Main Authors: Marc Ruoß, Georg Damm, Massoud Vosough, Lisa Ehret, Carl Grom-Baumgarten, Martin Petkov, Silvio Naddalin, Ruth Ladurner, Daniel Seehofer, Andreas Nussler, Sahar Sajadian
Format: Article
Language:English
Published: MDPI AG 2019-01-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:http://www.mdpi.com/1422-0067/20/2/347
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spelling doaj-88bc65d48ec24517b52114e1aaf394ab2020-11-25T01:48:38ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-01-0120234710.3390/ijms20020347ijms20020347Epigenetic Modifications of the Liver Tumor Cell Line HepG2 Increase Their Drug Metabolic CapacityMarc Ruoß0Georg Damm1Massoud Vosough2Lisa Ehret3Carl Grom-Baumgarten4Martin Petkov5Silvio Naddalin6Ruth Ladurner7Daniel Seehofer8Andreas Nussler9Sahar Sajadian10Siegfried Weller Institute, BG Trauma Clinic, Eberhard Karls University Tübingen, 72076 Tübingen, GermanyDepartment of Hepatobiliary Surgery and Visceral Transplantation, University of Leipzig, 04103 Leipzig, GermanyRoyan Institute for Stem Cell Biology and Technology, Department of Stem Cells and Developmental Biology, Tehran 16635-148, IranSiegfried Weller Institute, BG Trauma Clinic, Eberhard Karls University Tübingen, 72076 Tübingen, GermanySiegfried Weller Institute, BG Trauma Clinic, Eberhard Karls University Tübingen, 72076 Tübingen, GermanySiegfried Weller Institute, BG Trauma Clinic, Eberhard Karls University Tübingen, 72076 Tübingen, GermanyDepartment of General, Visceral and Transplant Surgery, University Hospital Tübingen, 72076 Tübingen, GermanyDepartment of General, Visceral and Transplant Surgery, University Hospital Tübingen, 72076 Tübingen, GermanyDepartment of Hepatobiliary Surgery and Visceral Transplantation, University of Leipzig, 04103 Leipzig, GermanySiegfried Weller Institute, BG Trauma Clinic, Eberhard Karls University Tübingen, 72076 Tübingen, GermanySiegfried Weller Institute, BG Trauma Clinic, Eberhard Karls University Tübingen, 72076 Tübingen, GermanyAlthough 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.http://www.mdpi.com/1422-0067/20/2/347tumor cellsepigenetic reprogrammingdrug metabolismhepatoma cell linesprimary human hepatocytes
collection DOAJ
language English
format Article
sources DOAJ
author Marc Ruoß
Georg Damm
Massoud Vosough
Lisa Ehret
Carl Grom-Baumgarten
Martin Petkov
Silvio Naddalin
Ruth Ladurner
Daniel Seehofer
Andreas Nussler
Sahar Sajadian
spellingShingle Marc Ruoß
Georg Damm
Massoud Vosough
Lisa Ehret
Carl Grom-Baumgarten
Martin Petkov
Silvio Naddalin
Ruth Ladurner
Daniel Seehofer
Andreas Nussler
Sahar Sajadian
Epigenetic Modifications of the Liver Tumor Cell Line HepG2 Increase Their Drug Metabolic Capacity
International Journal of Molecular Sciences
tumor cells
epigenetic reprogramming
drug metabolism
hepatoma cell lines
primary human hepatocytes
author_facet Marc Ruoß
Georg Damm
Massoud Vosough
Lisa Ehret
Carl Grom-Baumgarten
Martin Petkov
Silvio Naddalin
Ruth Ladurner
Daniel Seehofer
Andreas Nussler
Sahar Sajadian
author_sort Marc Ruoß
title Epigenetic Modifications of the Liver Tumor Cell Line HepG2 Increase Their Drug Metabolic Capacity
title_short Epigenetic Modifications of the Liver Tumor Cell Line HepG2 Increase Their Drug Metabolic Capacity
title_full Epigenetic Modifications of the Liver Tumor Cell Line HepG2 Increase Their Drug Metabolic Capacity
title_fullStr Epigenetic Modifications of the Liver Tumor Cell Line HepG2 Increase Their Drug Metabolic Capacity
title_full_unstemmed Epigenetic Modifications of the Liver Tumor Cell Line HepG2 Increase Their Drug Metabolic Capacity
title_sort epigenetic modifications of the liver tumor cell line hepg2 increase their drug metabolic capacity
publisher MDPI AG
series International Journal of Molecular Sciences
issn 1422-0067
publishDate 2019-01-01
description 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.
topic tumor cells
epigenetic reprogramming
drug metabolism
hepatoma cell lines
primary human hepatocytes
url http://www.mdpi.com/1422-0067/20/2/347
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