Comparison of the metabolic activation of environmental carcinogens in mouse embryonic stem cells and mouse embryonic fibroblasts
(2015) In Toxicology in Vitro 29(1). p.34-43- Abstract
We compared mouse embryonic stem (ES) cells and fibroblasts (MEFs) for their ability to metabolically activate the environmental carcinogens benzo[a]pyrene (BaP), 3-nitrobenzanthrone (3-NBA) and aristolochic acid I (AAI), measuring DNA adduct formation by (32)P-postlabelling and expression of xenobiotic-metabolism genes by quantitative real-time PCR. At 2 μM, BaP induced Cyp1a1 expression in MEFs to a much greater extent than in ES cells and formed 45 times more adducts. Nqo1 mRNA expression was increased by 3-NBA in both cell types but induction was higher in MEFs, as was adduct formation. For AAI, DNA binding was over 450 times higher in MEFs than in ES cells, although Nqo1 and Cyp1a1 transcriptional levels did not explain this... (More)
We compared mouse embryonic stem (ES) cells and fibroblasts (MEFs) for their ability to metabolically activate the environmental carcinogens benzo[a]pyrene (BaP), 3-nitrobenzanthrone (3-NBA) and aristolochic acid I (AAI), measuring DNA adduct formation by (32)P-postlabelling and expression of xenobiotic-metabolism genes by quantitative real-time PCR. At 2 μM, BaP induced Cyp1a1 expression in MEFs to a much greater extent than in ES cells and formed 45 times more adducts. Nqo1 mRNA expression was increased by 3-NBA in both cell types but induction was higher in MEFs, as was adduct formation. For AAI, DNA binding was over 450 times higher in MEFs than in ES cells, although Nqo1 and Cyp1a1 transcriptional levels did not explain this difference. We found higher global methylation of DNA in ES cells than in MEFs, which suggests higher chromatin density and lower accessibility of the DNA to DNA damaging agents in ES cells. However, AAI treatment did not alter DNA methylation. Thus mouse ES cells and MEFs have the metabolic competence to activate a number of environmental carcinogens, but MEFs have lower global DNA methylation and higher metabolic capacity than mouse ES cells.
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- author
- Krais, Annette M LU ; Mühlbauer, Karl-Rudolf ; Kucab, Jill E ; Chinbuah, Helena ; Cornelius, Michael G ; Wei, Quan-Xiang ; Hollstein, Monica ; Phillips, David H. ; Arlt, Volker M and Schmeiser, Heinz H.
- publishing date
- 2015-02
- type
- Contribution to journal
- publication status
- published
- keywords
- Animals, Aristolochic Acids, Benz(a)Anthracenes, Benzo(a)pyrene, Blotting, Western, Carcinogens, Environmental, DNA Adducts, DNA Methylation, Embryonic Stem Cells, Fibroblasts, Gene Expression Profiling, Mice, Mice, Inbred C57BL, Real-Time Polymerase Chain Reaction, Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
- in
- Toxicology in Vitro
- volume
- 29
- issue
- 1
- pages
- 10 pages
- publisher
- Elsevier
- external identifiers
-
- scopus:84930982021
- pmid:25230394
- ISSN
- 1879-3177
- DOI
- 10.1016/j.tiv.2014.09.004
- language
- English
- LU publication?
- no
- id
- b7596b24-0490-4807-b330-2ef5cb6a7e34
- date added to LUP
- 2017-10-17 15:29:39
- date last changed
- 2024-10-14 15:20:39
@article{b7596b24-0490-4807-b330-2ef5cb6a7e34, abstract = {{<p>We compared mouse embryonic stem (ES) cells and fibroblasts (MEFs) for their ability to metabolically activate the environmental carcinogens benzo[a]pyrene (BaP), 3-nitrobenzanthrone (3-NBA) and aristolochic acid I (AAI), measuring DNA adduct formation by (32)P-postlabelling and expression of xenobiotic-metabolism genes by quantitative real-time PCR. At 2 μM, BaP induced Cyp1a1 expression in MEFs to a much greater extent than in ES cells and formed 45 times more adducts. Nqo1 mRNA expression was increased by 3-NBA in both cell types but induction was higher in MEFs, as was adduct formation. For AAI, DNA binding was over 450 times higher in MEFs than in ES cells, although Nqo1 and Cyp1a1 transcriptional levels did not explain this difference. We found higher global methylation of DNA in ES cells than in MEFs, which suggests higher chromatin density and lower accessibility of the DNA to DNA damaging agents in ES cells. However, AAI treatment did not alter DNA methylation. Thus mouse ES cells and MEFs have the metabolic competence to activate a number of environmental carcinogens, but MEFs have lower global DNA methylation and higher metabolic capacity than mouse ES cells.</p>}}, author = {{Krais, Annette M and Mühlbauer, Karl-Rudolf and Kucab, Jill E and Chinbuah, Helena and Cornelius, Michael G and Wei, Quan-Xiang and Hollstein, Monica and Phillips, David H. and Arlt, Volker M and Schmeiser, Heinz H.}}, issn = {{1879-3177}}, keywords = {{Animals; Aristolochic Acids; Benz(a)Anthracenes; Benzo(a)pyrene; Blotting, Western; Carcinogens, Environmental; DNA Adducts; DNA Methylation; Embryonic Stem Cells; Fibroblasts; Gene Expression Profiling; Mice; Mice, Inbred C57BL; Real-Time Polymerase Chain Reaction; Comparative Study; Journal Article; Research Support, Non-U.S. Gov't}}, language = {{eng}}, number = {{1}}, pages = {{34--43}}, publisher = {{Elsevier}}, series = {{Toxicology in Vitro}}, title = {{Comparison of the metabolic activation of environmental carcinogens in mouse embryonic stem cells and mouse embryonic fibroblasts}}, url = {{http://dx.doi.org/10.1016/j.tiv.2014.09.004}}, doi = {{10.1016/j.tiv.2014.09.004}}, volume = {{29}}, year = {{2015}}, }