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@INPROCEEDINGS{Knops:839900,
author = {Knops, Katja and Boldt, Sonja and Wolkenhauer, Olaf and
Kriehuber, Ralf},
title = {{RADIATION}-{SPECIFIC} {GENE} {EXPRESSION} {CHANGES} {IN}
{HUMAN} {PBL} {AFTER} {EX} {VIVO} {IRRADIATION} {SUITABLE}
{FOR} {RADIATION} {BIODOSIMETRY}},
reportid = {FZJ-2017-07478},
year = {2011},
abstract = {Introduction: In case of a large-scale radiation accident
with involvement of individuals without physical dosimeters
it is important to identify individuals who have received a
moderate to high radiation dose to ensure proper medical
care. As current methods are time-consuming, a fast and
reliable method based on gene expression alterations is
developed.Methods: Human blood of 3 male and 3 female
healthy donors, belonging to 3 different age classes, was
irradiated ex vivo with 0, 0.02, 0.1, 0.5, 1, 2 and 4 Gy
(γ-rays, Cs-137). Peripheral blood lymphocytes (PBL) were
isolated and cultured for 6, 24 and 48 h in the medium- and
high dose range (0.5 – 4 Gy) and for 24 and 48 h after low
dose irradiation (0.02 and 0.1 Gy). At these times RNA and
proteins were isolated and RNA was applied for processing
whole human genome DNA-microarrays to analyse expression
profiles. In the medium- and high dose range the most robust
altered genes were selected for further qRT-PCR and protein
expression analysis. To examine the radiation-specificity of
the candidate genes, PBL were exposed to the DNA-damaging
agents Paracetamol (25 and 200 µg/ml) and Mitomycin C (0.1
and 0.4 µg/ml) for 6, 24 and 48 h and gene expression was
accordingly analyzed.Results: By a p-value and fold-change
driven gene selection 9 genes were identified in the low
dose range and 16 genes in the medium- and high dose range
allowing a radiation dose prediction accuracy of $96\%$
independently on the time-point post irradiation up to 48 h.
For 6 predictive genes in the medium- and high dose range
qRT-PCR measurements based on pooled and non-pooled
irradiated samples additionally validated the observed
radiation-induced gene expression alterations. Furthermore,
qRT-PCR analysis showed that the strong up-regulation of
these genes is highly radiation-specific as the
up-regulation after irradiation was much more pronounced
when compared to exposure with Paracetamol or Mitomycin C.
Protein expression analysis showed only for two genes a weak
correlation between gene and protein expression after
irradiation. Conclusion: In vitro gene expression analysis
in human PBL based on whole human DNA-microarray data
allowed identifying a rather small set of radiation dose
predictive and radiation-specific genes with a high
potential for biodosimetric applications in vivo after low-,
medium and high dose exposure. Funded by Kompetenzverbund
Strahlenforschung (KVSF), Bundesministerium für Bildung und
Forschung (BMBF), Project No.: 02NUK005A and 02NUK005B},
month = {Sep},
date = {2011-09-13},
organization = {14. Jahrestagung der Gesellschaft für
Biologische Strahlenforschung,
Rheinbach (Germany), 13 Sep 2011 - 16
Sep 2011},
subtyp = {After Call},
cin = {S-US},
cid = {I:(DE-Juel1)S-US-20090406},
pnm = {899 - ohne Topic (POF3-899)},
pid = {G:(DE-HGF)POF3-899},
typ = {PUB:(DE-HGF)24},
url = {https://juser.fz-juelich.de/record/839900},
}