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Cadmium, cobalt and lead cause stress response, cell cycle deregulation and increased steroid as well as xenobiotic metabolism in primary normal human bronchial epithelial cells which is coordinated by at least nine transcription factors

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Abstract

Workers occupationally exposed to cadmium, cobalt and lead have been reported to have increased levels of DNA damage. To analyze whether in vivo relevant concentrations of heavy metals cause systematic alterations in RNA expression patterns, we performed a gene array study using primary normal human bronchial epithelial cells. Cells were incubated with 15 μg/l Cd(II), 25 μg/l Co(II) and 550 μg/l Pb(II) either with individual substances or in combination. Differentially expressed genes were filtered out and used to identify enriched GO categories as well as KEGG pathways and to identify transcription factors whose binding sites are enriched in a given set of promoters. Interestingly, combined exposure to Cd(II), Co(II) and Pb(II) caused a coordinated response of at least seven stress response-related transcription factors, namely Oct-1, HIC1, TGIF, CREB, ATF4, SRF and YY1. A stress response was further corroborated by up regulation of genes involved in glutathione metabolism. A second major response to heavy metal exposure was deregulation of the cell cycle as evidenced by down regulation of the transcription factors ELK-1 and the Ets transcription factor GABP, as well as deregulation of genes involved in purine and pyrimidine metabolism. A third and surprising response was up regulation of genes involved in steroid metabolism, whereby promoter analysis identified up regulation of SRY that is known to play a role in sex determination. A forth response was up regulation of xenobiotic metabolising enzymes, particularly of dihydrodiol dehydrogenases 1 and 2 (AKR1C1, AKR1C2). Incubations with individual heavy metals showed that the response of AKR1C1 and AKR1C2 was predominantly caused by lead. In conclusion, we have shown that in vivo relevant concentrations of Cd(II), Co(II) and Pb(II) cause a complex and coordinated response in normal human bronchial epithelial cells. This study gives an overview of the most responsive genes.

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Acknowledgments

Microarray analysis was conducted at the IZKF Leipzig at the Faculty of Medicine of the University Leipzig (Projekt Z03).

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Authors and Affiliations

  1. Institute of Environmental Toxicology, Martin-Luther-University, Halle-Wittenberg, Franzosenweg 1a, 06097, Halle/Saale, Germany

    Felix Glahn, Jan Wiese & Heidi Foth

  2. Leibniz Institute for Natural Product Research and Infection Biology, Hans Knoell Institute, Jena, Germany

    Wolfgang Schmidt-Heck & Reinhard Guthke

  3. Institute of Biochemistry, Medical Faculty, University of Leipzig, Leipzig, Germany

    Sebastian Zellmer & Rolf Gebhardt

  4. IfADo, Leibniz Research Centre for Working Environment and Human Factors, Ardeystraße 67, 44139, Dortmund, Germany

    Klaus Golka, Gisela H. Degen, Matthias Hermes, Wiebke Schormann, Marc Brulport, Alexander Bauer, Essam Bedawy, Jan G. Hengstler & Jan G. Hengstler

  5. ISAS, Institute for Analytical Sciences, Bunsen-Kirchhoff-Str. 11, 44139, Dortmund, Germany

    Roland Hergenröder

  6. Translational Centre for Regenerative Medicine, 04103, Leipzig, Germany

    Thomas Lehmann

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204_2008_331_MOESM2_ESM.doc

Supplemental Table 1: Influence of incubation with a combination of Cd(II), Co(II) and Pb(II) (0.28, 0.29 and 3.11 mg/l, respectively) on gene expression patterns in bronchial epithelial cells. Kegg pathways, biological processes, molecular functions and cellular components are given that are up or downregulated in cells of at least three of the four patients. (DOC 216 kb)

204_2008_331_MOESM3_ESM.xls

Supplemental Table 3A: Genes that are at least 2-fold up regulated in bronchial epithelial cells only after incubation with 550 µg/l Pb(II) but are not up regulated after incubation with 15 µg/l Cd(II) or 25 µg/l Co(II).Supplemental Table 3B: Genes that are at least 2-fold up regulated in bronchial epithelial cells only after incubation with 15 µg/l Cd(II) but are not up regulated after incubation with or 25 µg/l Co(II) or with 550 µg/l Pb(II). No genes could be identified are at least 2-fold up regulated after incubation with 15 µg/l Co(II) but are not deregulated after incubation with or 25 µg/l Co(II) or with 550 µg/l Pb(II).No genes could be identified that are (XLS 5085 kb)

204_2008_331_MOESM4_ESM.xls

Supplemental Table 3C: Genes that are deregulated only after combined incubation with 15 µg/l Cd(II), 25 µg/l Co(II) and 550 µg/l Pb(II) but are not altered after individual incubations with either 15 µg/l Cd(II), or 25 µg/l Co(II) or 550 µg/l Pb(II) (XLS 18 kb)

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Glahn, F., Schmidt-Heck, W., Zellmer, S. et al. Cadmium, cobalt and lead cause stress response, cell cycle deregulation and increased steroid as well as xenobiotic metabolism in primary normal human bronchial epithelial cells which is coordinated by at least nine transcription factors. Arch Toxicol 82, 513–524 (2008). https://doi.org/10.1007/s00204-008-0331-9

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