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Integrative proteomic and microRNA analysis of primary human coronary artery endothelial cells exposed to low-dose gamma radiation

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Abstract

High doses of ionising radiation significantly increase the risk of cardiovascular disease (CVD), the vascular endothelium representing one of the main targets. Whether radiation doses lower than 500 mGy induce cardiovascular damage is controversial. The aim of this study was to investigate radiation-induced expression changes on protein and microRNA (miRNA) level in primary human coronary artery endothelial cells after a single 200 mGy radiation dose (Co-60). Using a multiplex gel-based proteomics technology (2D-DIGE), we identified 28 deregulated proteins showing more than ±1.5-fold expression change in comparison with non-exposed cells. A great majority of the proteins showed up-regulation. Bioinformatics analysis indicated “cellular assembly and organisation, cellular function and maintenance and molecular transport” as the most significant radiation-responsive network. Caspase-3, a central regulator of this network, was confirmed to be up-regulated using immunoblotting. We also analysed radiation-induced alterations in the level of six miRNAs known to play a role either in CVD or in radiation response. The expression of miR-21 and miR-146b showed significant radiation-induced deregulation. Using miRNA target prediction, three proteins found differentially expressed in this study were identified as putative candidates for miR-21 regulation. A negative correlation was observed between miR-21 levels and the predicted target proteins, desmoglein 1, phosphoglucomutase and target of Myb protein. This study shows for the first time that a low-dose exposure has a significant impact on miRNA expression that is directly related to protein expression alterations. The data presented here may facilitate the discovery of low-dose biomarkers of radiation-induced cardiovascular damage.

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Abbreviations

IR:

Ionising radiation

Gy:

Gray

miRNA:

microRNA

HCAEC:

Human coronary artery endothelial cells

IPA:

Ingenuity pathway analysis

DIGE:

Difference gel electrophoresis

CVD:

Cardiovascular disease

sqrt:

Square root

mirSVR:

miRNA score threshold

MS:

Mass spectrometry

2D-DIGE:

Two-dimensional difference gel electrophoresis

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Acknowledgments

The research leading to these results is supported by a grant from the European Community’s Seventh Framework Programme (EURATOM) contract no. 211403 (CARDIORISK). We thank Prof. Wolfgang Heidenreich for valuable advice in statistical questions, and Sandra Helm, Emily Hsu and Stefanie Winkler for technical assistance.

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Authors and Affiliations

  1. Institute of Radiation Biology, Helmholtz Zentrum München, German Research Center for Environmental Health, Ingolstaedter Landstrasse 1, 85764, Neuherberg, Germany

    Zarko Barjaktarovic, Natasa Anastasov, Omid Azimzadeh, Arundhathi Sriharshan, Michael J. Atkinson & Soile Tapio

  2. STUK—Radiation and Nuclear Safety Authority, Helsinki, Finland

    Hanna Tammio, Arvi Hakanen & Dariusz Leszczynski

  3. Department of Protein Science, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Germany

    Hakan Sarioglu & Marius Ueffing

  4. Research Unit of Radiation Cytogenetics, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Germany

    Arundhathi Sriharshan

Authors
  1. Zarko Barjaktarovic
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  2. Natasa Anastasov
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  3. Omid Azimzadeh
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  4. Arundhathi Sriharshan
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  5. Hakan Sarioglu
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  6. Marius Ueffing
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  7. Hanna Tammio
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  8. Arvi Hakanen
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  9. Dariusz Leszczynski
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  10. Michael J. Atkinson
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  11. Soile Tapio
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Correspondence to Zarko Barjaktarovic.

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Barjaktarovic, Z., Anastasov, N., Azimzadeh, O. et al. Integrative proteomic and microRNA analysis of primary human coronary artery endothelial cells exposed to low-dose gamma radiation. Radiat Environ Biophys 52, 87–98 (2013). https://doi.org/10.1007/s00411-012-0439-4

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