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Quantitative comparison of in vitro genotoxicity between metabolically competent HepaRG cells and HepG2 cells using the high-throughput high-content CometChip assay

  • Genotoxicity and Carcinogenicity
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Abstract

In vitro genotoxicity testing that employs metabolically active human cells may be better suited for evaluating human in vivo genotoxicity than current bacterial or non-metabolically active mammalian cell systems. In the current study, 28 compounds, known to have different genotoxicity and carcinogenicity modes of action (MoAs), were evaluated over a wide range of concentrations for the ability to induce DNA damage in human HepG2 and HepaRG cells. DNA damage dose–responses in both cell lines were quantified using a combination of high-throughput high-content (HTHC) CometChip technology and benchmark dose (BMD) quantitative approaches. Assays of metabolic activity indicated that differentiated HepaRG cells had much higher levels of cytochromes P450 activity than did HepG2 cells. DNA damage was observed for four and two out of five indirect-acting genotoxic carcinogens in HepaRG and HepG2 cells, respectively. Four out of seven direct-acting carcinogens were positive in both cell lines, with two of the three negatives being genotoxic mainly through aneugenicity. The four chemicals positive in both cell lines generated HTHC Comet data in HepaRG and HepG2 cells with comparable BMD values. All the non-genotoxic compounds, including six non-genotoxic carcinogens, were negative in HepaRG cells; five genotoxic non-carcinogens also were negative. Our results indicate that the HTHC CometChip assay detects a greater proportion of genotoxic carcinogens requiring metabolic activation (i.e., indirect carcinogens) when conducted with HepaRG cells than with HepG2 cells. In addition, BMD genotoxicity potency estimate is useful for quantitatively evaluating CometChip assay data in a scientifically rigorous manner.

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Acknowledgements

J.E.S. and K.D. were supported by appointments to the Postgraduate Research Program at the National Center for Toxicological Research (NCTR) administered by the Oak Ridge Institute for Science Education through an interagency agreement between the U.S. Department of Energy and the U.S. Food and Drug Administration (FDA). We greatly appreciate Dr. Lei Guo (DBT/NCTR) for generously providing HepG2 cells and thank Drs. Robert H. Heflich, Dayton Petibone, and Elvis-Yane Cuevas-Martinez for their critical review of this article.

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  1. Kostiantyn Dreval

    Present address: Department of Internal Medicine, Division of Molecular Medicine, Program in Cancer Genetics, Epigenetics and Genomics, University of New Mexico Comprehensive Cancer Center, Albuquerque, NM, 87131, USA

Authors and Affiliations

  1. Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, Jefferson, AR, 72079, USA

    Ji-Eun Seo, Nan Mei & Xiaoqing Guo

  2. Division of Biochemical Toxicology, National Center for Toxicological Research, Jefferson, AR, 72079, USA

    Volodymyr Tryndyak, Qiangen Wu, Kostiantyn Dreval, Igor Pogribny & Matthew Bryant

  3. Center for Veterinary Medicine, U.S. Food and Drug Administration, Rockville, MD, 20855, USA

    Tong Zhou

  4. Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, 20993, USA

    Timothy W. Robison

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Correspondence to Xiaoqing Guo.

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Seo, JE., Tryndyak, V., Wu, Q. et al. Quantitative comparison of in vitro genotoxicity between metabolically competent HepaRG cells and HepG2 cells using the high-throughput high-content CometChip assay. Arch Toxicol 93, 1433–1448 (2019). https://doi.org/10.1007/s00204-019-02406-9

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