Archives of Toxicology

, Volume 87, Issue 3, pp 505–515 | Cite as

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

  • P. C. E. van Kesteren
  • P. E. Zwart
  • M. M. Schaap
  • T. E. Pronk
  • M. H. M. van Herwijnen
  • J. C. S. Kleinjans
  • B. G. H. Bokkers
  • R. W. L. Godschalk
  • M. J. Zeilmaker
  • H. van Steeg
  • M. Luijten
In vitro systems

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.

Keywords

Toxicogenomics Carcinogenesis Benzo[a]pyrene Xpa−/−p53+/− Physiologically based kinetic modeling 

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+/−

Supplementary material

204_2012_949_MOESM1_ESM.pdf (47 kb)
Supplementary material 1 (PDF 47 kb)
204_2012_949_MOESM2_ESM.pdf (45 kb)
Supplementary material 2 (PDF 45 kb)
204_2012_949_MOESM3_ESM.pdf (21 kb)
Supplementary material 3 (PDF 21 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • P. C. E. van Kesteren
    • 1
    • 2
  • P. E. Zwart
    • 1
  • M. M. Schaap
    • 1
    • 3
  • T. E. Pronk
    • 1
    • 2
  • M. H. M. van Herwijnen
    • 2
  • J. C. S. Kleinjans
    • 2
  • B. G. H. Bokkers
    • 4
  • R. W. L. Godschalk
    • 5
  • M. J. Zeilmaker
    • 4
  • H. van Steeg
    • 1
    • 3
  • M. Luijten
    • 1
  1. 1.Laboratory for Health Protection ResearchNational Institute for Public Health and the EnvironmentBilthovenThe Netherlands
  2. 2.Department of ToxicogenomicsMaastricht UniversityMaastrichtThe Netherlands
  3. 3.Department of ToxicogeneticsLeiden University Medical Center (LUMC)LeidenThe Netherlands
  4. 4.Centre for Substances and Integral Risk AssessmentNational Institute for Public Health and the Environment (RIVM)BilthovenThe Netherlands
  5. 5.Department of Toxicology, School for Nutrition, Toxicology and Metabolism (NUTRIM)Maastricht UniversityMaastrichtThe Netherlands

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