Cited by 44Pharmacogenetics and Drug MetabolismDrug-Induced Hepatotoxicity and ProtectionDrug Transport and Resistance Mechanisms

Characterization of cytochrome P450s (CYP)-overexpressing HepG2 cells for assessing drug and chemical-induced liver toxicity

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Chen, Si, Wu, Qiangen, Li, Xilin, Li, Dongying, Mei, Nan, Ning, Baitang, Puig, Montserrat, Ren, Zhen +2 more·2021·Journal of Environmental Science and Health, Part C

Abstract

Hepatic metabolism catalyzed by the cytochrome P450 (CYP) superfamily affects liver toxicity associated with exposures to natural compounds and xenobiotic agents. Previously we generated a battery of HepG2-derived stable cell lines that individually express 14 CYPs (1A1, 1A2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, 3A4, 3A5, and 3A7). In this study, we comprehensively characterized each cell line for its CYP expression and enzyme activity. Specifically, we measured the mRNA expression, protein expression, and metabolite formation. Using CYP3A4, 2D6, and 2C9-overexpressing cells as representatives, we examined the stability of these cells in long-term cultures for up to 10 passages. The results showed that CYPs can be stably overexpressed for up to 10 cell culture passages without losing their activities. The robustness of responses to stimuli among the cells at different passages was also investigated in CYP3A4-overexpressing cells and the response to amiodarone and dronedarone showed no difference between the cells at the passage 2 and 10. Moreover, the mRNA expression level of most CYPs was higher in CYP-overexpressing HepG2 cells than that in HepaRG cells and primary human hepatocytes. This study confirmed the stability of CYP-overexpressing HepG2 cell lines and provided useful information for a broader use of these cells in pharmacologic and toxicologic research.

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Crossref · 10.1080/26896583.2021.1880242

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  • HepG2 is suitable for studying general mechanisms of cytotoxicity and cellular stress response but limited for quantitative prediction of metabolism and biotransformation of many compounds.

    “Recent reviews and experimental studies directly interpret HepG2 as a model suitable for solving certain toxicological problems, in particular for studying general mechanisms of cytotoxicity and cellular stress response (e.g., activation of transcription factors activator protein-1 (AP1), p53, nuclear factor-erythroid 2 related factor 2 (Nrf2), and nuclear factor kappa B (NF-κB)) [109,110,111],”

    From The HepG2 Cell Line as a Model for Studying Metabolic Dysfunction-Associated Steatotic Liver Disease (2026) · cites this paper as [113]

    The Introduction describes HepG2 as frequently used to elucidate mechanisms of toxicity and says its reduced drug-metabolizing enzyme expression limits metabolism-related studies. The Discussion likewise notes limited drug metabolic capacity in parental HepG2 cells. Our copy does not establish the cited sentence’s more specific claim about studying cellular stress responses through AP1, p53, Nrf2, or NF-κB activation. (LLM, conf=0.84)

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  • Stable HepG2 panels with overexpression of individual CYPs have demonstrated suitability for identifying metabolically mediated toxicity and for determining the contribution of a specific isoform to the biotransformation of a xenobiotic.

    “Stable HepG2 panels with overexpression of individual CYPs have already demonstrated their suitability for identifying metabolically mediated toxicity and for determining the contribution of a specific isoform to the biotransformation of a xenobiotic [112,113].”

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Authors
Chen, S., Wu, Q., Li, X., Li, D., Mei, N., Ning, B., Puig, M., Ren, Z., Tolleson, W. H., & Guo, L.
Affiliations
  • National Center for Toxicological Research (US)
  • United States Food and Drug Administration (US)
Corresponding author
Guo, Lei
Venue
Journal of Environmental Science and Health, Part C
Published
2021-01-02
Volume
39
Issue
1
Pages
68-86
ISSN
  • 1059-0501 (ISSN-L)
  • 2689-6583
  • 2689-6591
OpenAlex
W3126287748
Cited by
44
References
65
Citations by year
2026: 5 · 2025: 13 · 2024: 9 · 2023: 6 · 2022: 6 · 2021: 5
FWCI
5.95
Type
journal article
Language
en
Open access
Locations
  • Journal of Environmental Science and Health Part C (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
National Institutes of Health (FD999999)
Topics
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
Keywords
cytochrome P450, stable cell lines, drug-induced liver injury, CYP3A4, CYP2D6, CYP2C9, CYP3A5, CYP3A7, CYP2C19, CYP2C18, CYP2C8, CYP2E1, CYP2A6, CYP2B6, cell line stability, amiodarone, dronedarone, hepatotoxicity assessment
MeSH terms
Dronedarone, Cell Line, Cytochrome P-450 Enzyme System, Pharmaceutical Preparations, Humans, Liver, Metabolic Clearance Rate, Inactivation, Metabolic, Microsomes, Liver, Oxidation-Reduction, Hepatocytes, Cytochrome P-450 CYP3A, Chemical and Drug Induced Liver Injury, Hep G2 Cells

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