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Comparative analysis of phase I and II enzyme activities in 5 hepatic cell lines identifies Huh-7 and HCC-T cells with the highest potential to study drug metabolism

  • Molecular Toxicology
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Abstract

Primary human hepatocytes (hHeps) are still gold standard to perform human drug metabolism studies, but their availability is limited by donor organ scarcity. Therefore, hepatoma cell lines are widely used as alternatives, although their phases I and II drug-metabolizing activities are substantially lower compared with hHeps. The major advantage of these cell lines is immediate availability, standardized culture conditions and unlimited life span. Therefore, the aim of this study was to investigate the drug-metabolizing profile of five human hepatoma cell lines (HepG2, Hep3B, HCC-T, HCC-M and Huh-7) over a culture period of 10 passages. Fluorescent-based assays for seven different cytochrome P450 (CYP) isoforms and seven different phase II enzymes were performed and compared with enzymatic activities of hHeps. CYP activities were much lower in the cell lines (5–15% of hHeps), whereas phase II enzyme activities that are involved in buffering oxidative stress (e.g., Glutathione-S-transferase) reached levels comparable with hHeps. Furthermore, phases I and II enzyme activities in hepatoma cell lines vary strongly during culture time. Interestingly, the most constant results were obtained with Huh-7 cells. Huh-7 cells as well as HCC-T cells exhibited a drug-metabolizing profile closest to hHeps between passages two and four. Toxicity studies with Diclofenac, Paracetamol and Verapamil in both cell lines show dose–response characteristics and EC50 values similar to hHeps. Therefore, we propose that due to the more consistent results throughout the passages, Huh-7 could be an alternative system to the limitedly available hHeps and frequently used HepG2 cell line in the study of drug metabolism.

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Abbreviations

AMMC:

3-[2-(N,N-diethyl-N-methylammonium)ethyl]-7-methoxy-4-methyl-coumarin

AHMC:

3-[2-(N,N-diethylamino)ethyl]-7-hydroxy-4-methylcoumarin

BFC:

7-benzyloxy-4-trifluoromethylcoumarin

CEC:

3-cyano-7-ethoxycoumarin

EFC:

7-ethoxy-4-trifluoromethylcoumarin

7-EC:

7-ethoxycoumarin

HFC:

7-hydroxy-4-trifluoromethylcoumarin

hHeps:

Primary human hepatocytes

MCB:

Monochlorobimane

MFC:

7-methoxy-4-trifluoromethylcoumarin

4-MU:

4-methylumbelliferone

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Acknowledgments

The authors are grateful to Ms. Marina Unger for excellent technical assistance and to Mr. Fritz Seidl for editing. This work was partially supported by grants from the German Federal Ministry of Education and Research BMBF 01GG0732 (to AKN), 0315208E (to AKN).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Traumatology, Technische Universität München, Ismaninger Straße 22, 81675, Munich, Germany

    Jie Lin, Lilianna Schyschka, Ruben Mühl-Benninghaus, Jan Neumann, Ulrich Stöckle, Andreas K. Nussler & Sabrina Ehnert

  2. Shantou Centre for Disease Control and Prevention, Shantou, 515031, Guangdong, Peoples’ Republic of China

    Jie Lin

  3. Department of Nutrition and Food Hygiene, Tongji Medical School, HUST, 430030, Wuhan, Peoples’ Republic of China

    Liping Hao & Liegang Liu

  4. Department of Surgery, Clinic Neuperlach, Oskar-Maria-Graf-Ring 51, 81737, Munich, Germany

    Natascha Nussler

  5. Department of Medicine II, University Hospital Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1–3, 68167, Mannheim, Germany

    Steven Dooley

  6. Department of Traumatology, Eberhard Karls Universität Tübingen, Schnarrenbergstraße 95, 72076, Tübingen, Germany

    Ulrich Stöckle, Andreas K. Nussler & Sabrina Ehnert

Authors
  1. Jie Lin
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  2. Lilianna Schyschka
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  3. Ruben Mühl-Benninghaus
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  4. Jan Neumann
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  5. Liping Hao
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  6. Natascha Nussler
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  7. Steven Dooley
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  8. Liegang Liu
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  9. Ulrich Stöckle
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  10. Andreas K. Nussler
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  11. Sabrina Ehnert
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Corresponding author

Correspondence to Andreas K. Nussler.

Additional information

Jie Lin, Lilianna Schyschka, Andreas K Nussler and Sabrina Ehnert contributed equally to the publication.

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Lin, J., Schyschka, L., Mühl-Benninghaus, R. et al. Comparative analysis of phase I and II enzyme activities in 5 hepatic cell lines identifies Huh-7 and HCC-T cells with the highest potential to study drug metabolism. Arch Toxicol 86, 87–95 (2012). https://doi.org/10.1007/s00204-011-0733-y

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