Abstract
Purpose: Identification of early radiation response genes (ERG) in human lymphocytes after γ-irradiation by using the whole-human-genome DNA-microarrays and the evaluation of their possible role in rapid radiation biodosimetry by applying real-time quantitative polymerase chain reaction (RT-qPCR) methodology for validation in a small group of human individuals.Materials and methods: Whole blood from a healthy human donor was exposed at 37°C to 137Cs γ-radiations (absorbed dose: 1–4 Gy). Fifteen minutes following irradiation the lymphocytes were isolated from the blood (for 2 h at 20°C) and their gene expression was investigated using the DNA-microarrays. Subsequently, 14 genes were selected and validated using the TaqMan™ probes based upon the RT-qPCR assay within a group of 6 human donors.Results: A dose-related relative change in quantitative gene expression using the DNA-microarray assay was demonstrated in 24 of 102 genes. Up-regulation of expression was observed in 15 genes: CD69 (CD69 molecule), CDKN1A (cyclin-dependent kinase inhibitor 1A), EGR1 (early growth response 1), EGR4 (early growth response 4), FLJ35725 (chromosome 4 ORF 23), hCG2041177 (hCG – human Celera® Genome), hCG1643466.2, IFN-γ (interferon-γ), ISG20L (interferon stimulated exonuclease gene 20 kDa – like 1), c-JUN (jun oncogene), MDM2 (mouse double minute 2), MUC5B (mucine), PLK2 (polo-like kinase 2), RND1 (rho-family GTPase 1) and TNFSF9 (tumour necrosis factor superfamily member 9). Down-regulation of expression was found in the remaining nine genes: GRIK3 (glutamate receptor ionotropic kainate 3), hCG1985174, hCG1998530, hCG2038519, OCLN (occludin), RPL10A (ribosomal protein L10a), SERHL2 (serine hydrolase-like 2), SGK3 (serum/glucocorticoid regulated kinase 3) and STARD13 (START domain containing 13).Conclusion: A significant correlation between absorbed radiation dose and change in relative gene expression was particularly evident for EGR1, EGR4, IFN-γ, c-JUN and TNFSF9 (p ≤ 0.05). Results warrant the further investigation of these ERG as potential biodosimetric markers.
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