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Low-dose irradiation causes rapid alterations to the proteome of the human endothelial cell line EA.hy926

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Abstract

High doses of ionising radiation damage the heart by an as yet unknown mechanism. A concern for radiological protection is the recent epidemiological data indicating that doses as low as 100–500 mGy may induce cardiac damage. The aim of this study was to identify potential molecular targets and/or mechanisms involved in the pathogenesis of low-dose radiation-induced cardiovascular disease. The vascular endothelium plays a pivotal role in the regulation of cardiac function and is therefore a potential target tissue. We report here that low-dose radiation induced rapid and time-dependent changes in the cytoplasmic proteome of the human endothelial cell line EA.hy926. The proteomes were investigated at 4 and 24 h after irradiation at two different dose rates (Co-60 gamma ray total dose 200 mGy; 20 mGy/min and 190 mGy/min) using 2D-DIGE technology. Differentially expressed proteins were identified, after in-gel trypsin digestion, by MALDI-TOF/TOF tandem mass spectrometry, and peptide mass fingerprint analyses. We identified 15 significantly differentially expressed proteins, of which 10 were up-regulated and 5 down-regulated, with more than ± 1.5-fold difference compared with unexposed cells. Pathways influenced by the low-dose exposures included the Ran and RhoA pathways, fatty acid metabolism and stress response.

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Abbreviations

D-MEM:

Dulbecco’s Modified Eagle’s Medium

PMMA:

Poly(methyl methacrylate)

HAT:

Hypoxanthine Aminopterin Thymidine

SNP:

Single nucleotide polymorphism

Gy:

Gray

o/n:

Overnight

TFA:

Trifluoroacetic acid

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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 Dr. Ludwig Hieber and Dr. Herbert Braselmann for giving valuable advice in statistical questions.

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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

    Franka Pluder, Zarko Barjaktarovic, Omid Azimzadeh, Simone Mörtl, Anne Krämer, Sylvia Steininger, Michael J. Atkinson & Soile Tapio

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

    Hakan Sarioglu

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

    Dariusz Leszczynski, Reetta Nylund & Arvi Hakanen

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

    Arundhathi Sriharshan

  5. Department of Radiation Oncology, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany

    Michael J. Atkinson

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  1. Franka Pluder
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  2. Zarko Barjaktarovic
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Correspondence to Soile Tapio.

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Pluder, F., Barjaktarovic, Z., Azimzadeh, O. et al. Low-dose irradiation causes rapid alterations to the proteome of the human endothelial cell line EA.hy926. Radiat Environ Biophys 50, 155–166 (2011). https://doi.org/10.1007/s00411-010-0342-9

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