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Benzo[a]pyrene-induced transcriptomic responses in primary hepatocytes and in vivo liver: Toxicokinetics is essential for in vivo–in vitro comparisons

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Abstract

The traditional 2-year cancer bioassay needs replacement by more cost-effective and predictive tests. The use of toxicogenomics in an in vitro system may provide a more high-throughput method to investigate early alterations induced by carcinogens. Recently, the differential gene expression response in wild-type and cancer-prone Xpa −/− p53 +/− primary mouse hepatocytes after exposure to benzo[a]pyrene (B[a]P) revealed downregulation of cancer-related pathways in Xpa −/− p53 +/− hepatocytes only. Here, we investigated pathway regulation upon in vivo B[a]P exposure of wild-type and Xpa −/− p53 +/− mice. In vivo transcriptomics analysis revealed a limited gene expression response in mouse livers, but with a significant induction of DNA replication and apoptotic/anti-apoptotic cellular responses in Xpa −/− p53 +/− livers only. In order to be able to make a meaningful in vivo–in vitro comparison we estimated internal in vivo B[a]P concentrations using DNA adduct levels and physiologically based kinetic modeling. Based on these results, the in vitro concentration that corresponded best with the internal in vivo dose was chosen. Comparison of in vivo and in vitro data demonstrated similarities in transcriptomics response: xenobiotic metabolism, lipid metabolism and oxidative stress. However, we were unable to detect cancer-related pathways in either wild-type or Xpa −/− p53 +/− exposed livers, which were previously found to be induced by B[a]P in Xpa −/− p53 +/− primary hepatocytes. In conclusion, we showed parallels in gene expression responses between livers and primary hepatocytes upon exposure to equivalent concentrations of B[a]P. Furthermore, we recommend considering toxicokinetics when modeling a complex in vivo endpoint with in vitro models.

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Abbreviations

ANOVA:

Analysis of variance

B[a]P:

Benzo[a]pyrene

BPDE:

Benzo[a]pyrene-7,8-diol-9,10-epoxide

GI:

Gastrointestinal

FDR:

False discovery rate

GenMAPP:

Gene Map Annotator and Pathway Profiler

GO:

Gene Ontology

KEGG:

Kyoto Encyclopedia of Genes and Genomes

PBK:

Physiologically based kinetic

PCA:

Principal component analysis

WT:

Wild-type

Xpa:

Xeroderma pigmentosum A

Xpa/p53 :

Xpa −/− p53 +/−

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Acknowledgments

We thank R. Vlug, J. Bos, H. Strootman and T. van de Kuil for their support with the hepatocytes isolations. We also thank Prof. Dr. J. Hengstler for his advice on hepatocyte isolation and culturing. This work was supported by the Technology Foundation STW [grant MFA6809].

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Laboratory for Health Protection Research, National Institute for Public Health and the Environment, P.O. Box 1, 3720 BA, Bilthoven, The Netherlands

    P. C. E. van Kesteren, P. E. Zwart, M. M. Schaap, T. E. Pronk, H. van Steeg & M. Luijten

  2. Department of Toxicogenomics, Maastricht University, Maastricht, The Netherlands

    P. C. E. van Kesteren, T. E. Pronk, M. H. M. van Herwijnen & J. C. S. Kleinjans

  3. Department of Toxicogenetics, Leiden University Medical Center (LUMC), Leiden, The Netherlands

    M. M. Schaap & H. van Steeg

  4. Centre for Substances and Integral Risk Assessment, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands

    B. G. H. Bokkers & M. J. Zeilmaker

  5. Department of Toxicology, School for Nutrition, Toxicology and Metabolism (NUTRIM), Maastricht University, Maastricht, The Netherlands

    R. W. L. Godschalk

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  1. P. C. E. van Kesteren
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  2. P. E. Zwart
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  3. M. M. Schaap
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  4. T. E. Pronk
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  5. M. H. M. van Herwijnen
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  6. J. C. S. Kleinjans
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  7. B. G. H. Bokkers
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  8. R. W. L. Godschalk
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  9. M. J. Zeilmaker
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  10. H. van Steeg
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  11. M. Luijten
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Correspondence to M. Luijten.

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van Kesteren, P.C.E., Zwart, P.E., Schaap, M.M. et al. Benzo[a]pyrene-induced transcriptomic responses in primary hepatocytes and in vivo liver: Toxicokinetics is essential for in vivo–in vitro comparisons. Arch Toxicol 87, 505–515 (2013). https://doi.org/10.1007/s00204-012-0949-5

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