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Mitochondrial DNA copy number and hnRNP A2/B1 protein: Biomarkers for direct exposure of benzene

Authors

  • Ha-Young Eom,

    1. Department of Laboratory Medicine, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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    • contributed equally to this article.

  • Hye-Ran Kim,

    1. Center for Biomedical Human Resources at Chonnam National University, Gwangju, Korea
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    • contributed equally to this article.

  • Hwan-Young Kim,

    1. Department of Laboratory Medicine, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Dong-Kyun Han,

    1. Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Hee-Jo Baek,

    1. Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Jae-Hyuk Lee,

    1. Department of Pathology, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Jai Dong Moon,

    1. Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Jong-Hee Shin,

    1. Department of Laboratory Medicine, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Soon-Pal Suh,

    1. Department of Laboratory Medicine, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Dong-Wook Ryang,

    1. Department of Laboratory Medicine, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Hoon Kook,

    1. Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
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  • Myung-Geun Shin

    Corresponding author
    1. Department of Laboratory Medicine, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea
    • Department of Laboratory Medicine, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea.
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Abstract

The present study was performed to identify biomarkers for exposure of benzene in blood cells and hematopoietic tissues. Peripheral mononuclear cells, hematopoietic stem cells, and leukemia cell lines were cultured in RPMI 1640 media with the addition of 0, 1, and 10 mM of benzene. Hydrogen peroxide was measured using an enzyme immunoassay. Mitochondrial mass, membrane potential, and mitochondrial DNA (mtDNA) copy number were measured using MitoTracker Green/Red probes, and real-time polymerase chain reaction. In addition, two-dimensional gel electrophoresis and mass spectrometry matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) technology were performed to identify protein markers. The mitochondrial contents and membrane potentials were dramatically increased after three weeks of direct benzene exposure. The hydrogen peroxide level increased significantly after two weeks of treatment with benzene (4.4 ± 1.9 µM/mg protein) compared to the non-benzene treatment group (1.2 ± 1.0; p = 0.001). The mtDNA copy number gradually increased after exposure to benzene. Numerous protein markers showed significant aberrant expression after exposure to benzene. Among them, the heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1 was markedly decreased after exposure to benzene. Thus, increased mitochondrial mass, mtDNA copy number, and the hnRNP A2/B1 protein were biomarkers for benzene-related toxicity and hematotoxicity. Environ. Toxicol. Chem. 2011;30:2762–2770. © 2011 SETAC

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