research from animal testing to clinical experience

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Ann Ist Super Sanit 2009 | Vol. 45, No. 3: 272-277

Cell response to ionising radiation

analysed by gene expression patterns
Laurence Roy, Gatan Gruel and Aurlie Vaurijoux
Institut de Radioprotection et de Sret Nuclaire, Direction de la Radioprotection de lHomme,
Fontenay-aux-Roses, France
Summary. Following ionising radiation exposure of living cells several mechanisms are activated through
gene modulations. The measurement of these modifications can be done with QT-PCR and, since about
10 years, microarray technique. The latter approach has the advantage to allow a global monitoring of the
complex cellular responses to radiation-induced stress and has been proposed to be used for dose assessment. Even if some publications have identified sets of genes specific to given doses, and that some of the
genes have an induction proportional to the dose, a precise estimation of the received dose seems difficult
with gene expression, at least in the near future. Nevertherless, in vivo studies have shown that gene profiles
of individuals chronically exposed to a cumulative dose of more than 10 mSv are significantly modified.
This highlights the great potential of microarray approaches in the detection of low dose exposure.
Key words: microarray, ionising radiation, gene modulation.

Riassunto (Risposta cellulare alle radazioni ionizzanti analizzata con profili di espressione genica).
Lesposizione di cellule viventi alle radiazioni ionizzanti attiva diversi meccanismi attraverso lespressione
genica. La misura di queste modificazioni pu essere fatta con la QT-PCR e, da circa un decennio con la
tecnologia a microarray. Questo approccio ha il vantaggio di permettere un monitoraggio globale della
complessa risposta delle cellule in seguito allo stress dovuto alla radiazione ionizzante ed stato proposto
per permettere la valutazione della dose. Anche se alcune pubblicazioni hanno identificato che esistono
set di geni specifici alle date dosi e che alcuni geni hanno uninduzione proporzionale alla dose, una stima
precisa della dose sembra difficilmente raggiungibile. Comunque, studi in vivo hanno mostrato che profili
genici di individui cronicamente esposti a una dose accumulata maggiore di 10 mSv sono significativamente modificati.
Parole chiave: microarray, radiazioni ionizzanti, espressione genica.

Introduction
There are three major reasons responsible for the
currently observed increased hazard of accidental
exposure to ionising radiation. First, the increasing
need for radiation sources in numerous industrial
applications (food sterilization, construction, engineering etc.) leads to an increasing probability of irregular uses, storages or losses of sources. Secondly,
advances in nuclear medicine generate new protocols and tools more efficient but also much more
complex to carry out, and increase the risk of an
accidental overexposure. Finally, the possibility of
a terrorist attack using a radiological or nuclear device has to be taken into account. All these issues
could lead to the exposure of one to several thousand individuals not wearing dosimeters.
The dicentric assay is the current standard for radiation biodosimetry but its application in the particular situation of a mass casualty is limited due to
time constraint. Indeed, in addition to the time required for the stimulation of cell division, the manual dicentric scoring is very time consuming. In addi-

tion, the sensitivity of this technique is 0.1 Gy. This

highlights the need to establish new biomarkers and
approaches to biodosimetry in order to be able to assign the individuals with significant exposure to appropriate medical care. Many alternatives have been
proposed [1] among which gene expression profiling
seems particularly promising. Indeed, technological
and methodological advances in genomics led to the
potential discovery of many radiation-responsive biomarkers in peripheral blood cells [2-7]. Molecular
pathways involved in radiation-induced stress response have first been discovered through traditional biochemical approaches that have been used to
monitor the activation of a single gene. However, it
is evident that this approach becomes restrictive and
does not allow a global monitoring of the complex
response of cells following ionising radiation stress.
The advent of cDNA microarray makes it possible to
quantify the entire transcriptome in a single experiment. Many studies were published since the initial
development of cDNA microarray hybridisation.
The ability to compare the levels of thousands of

Address for correspondence: Laurence Roy, Laboratoire de Dosimtrie Biologique, Institut de Radioprotection et de Sret
Nuclaire, BP 17, F-92262 Fontenay-aux-Roses, France. E-mail: Laurence.roy@irsn.fr.

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Radiation dose assessment using microarray

mRNA simultaneously in a single hybridisation has

the potential to contribute to the understanding of
many different biological mechanisms. The purpose
of this review is to analyse to what degree the development of methods to detect gene expression allows
them to be applied in the field of biological dosimetry. To validate a biodosimeter, several parameters
shall be verified such as sensitivity, reproducibility,
sensitivity to dose-rate, radiation energy and linear
energy transfer response, as well as sources of variability and uncertainty. All biological methods are
subject to variability that are often beyond control
and represent a major source of uncertainty. These
include variations in background signals not directly
associated with radiation exposure, inter- and intraindividual variability of radiation response, and genetic and environmental effects [8]. The evaluation
of these parameters is not easy as it should be done
on individuals exposed in vivo to ionising radiation.
Some workers are exposed chronically to low levels
of ionising radiations but the regulation limits this
exposure to 20 mSv over one year. Therefore, the occupational doses do not allow to validate a new bioindicator over a large dose range. Higher doses can
be reached when individuals are accidental exposed
but such cases are rare and therefore most validations have to be performed on in vitro exposed human samples or animal models.
Extrapolation from one tissue to another is difficult, as mentioned by Zhao et al. [9], who compared gene expression in cells from mouse kidney
and brain exposed in vivo to ionising radiation. They
observed that cells of different origin respond to the
damaging effects of radiation in very different ways.
Similar results have been obtained by Mori et al. [10]
who compared the level of mRNA in freshly isolated
CD4+ T lymphocytes with CD4+ T lymphocytesderived Jurkat leukemic cells. Both cell types were
exposed to a dose corresponding to the lethal dose
50 (LD50 - 1 Gy for the isolated lymphocytes and
0.5 Gy for the Jutkat cells). Due to the difficulties
to extrapolate from one cell type to another we have
restricted our review to studies done on non-transformed human cells (cell lines not included). Both in
vivo and in vitro exposures were selected. Except in
some few cases, publications involving animal studies were not included in this review.
Methodologies used
Levels of mRNA transcript can be measured and
quantified using quantitative polymerase chain reaction (QT-PCR). This technique allows only the
quantification of few gene modulations at a time. A
review of the genes that are modulated in response
to ionising radiation is provided by Chaudhry [11].
Expression profiling with oligonucleotide microarrays has opened up new possibilities for large-scale
gene expression studies [12]. The use of this technology can lead to the characterization of biomarkers
for early diagnosis and prognosis, and may guide

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treatment and treatment innovation. This technology has been used successfully for tumour classification and for predicting the response of individual
tumours to specific treatment [13, 14]. It also seems
to be a useful method for detecting subtle changes in
transcription following exposure to ionising radiation [2, 15, 16]. The principle of this technique is not
described here but can be found in Amundson and
Fornace [17]. A major limitation of this approach
is that large numbers of repeat hybridizations are
required to ensure the reproducibility and statistical validity of the data. Microarray analysis raises
new challenges for statistical methodology, due to
the huge number of hypotheses tested in parallel.
Generally, too few samples are tested for estimation
of a real multivariate distribution of the expressed
genes. The main consequence is a high risk of selecting false positive signals. Several methods for
controlling the type I error rate (false positives) due
to multiple tests have been reviewed and adapted to
microarray technology. Two general strategies of
analysis (not exclusive) can be highlighted: the first
one consists of p-value correction methods [18].
This strategy is designed to identify genes and group
of genes involved in the cell response to ionising
radiation. The stringency of this methodology can
be modulated depending on the chosen p-value correction. Lowering the stringency of probe selection
increases the risk of false positives, but can allow to
identify consistent groups of genes (in terms of protein function, type of regulation, localisation etc.) in
order to contribute to the understanding of the cell
response to ionising radiation. The second strategy
is intended for identification of robust signature of
exposure. Theoretically, this strategy requires the
splitting of the data set in two independent parts:
one (learning set) for the isolation of the putative
signature of a condition (for example: exposed versus non-exposed), the second (challenge set) to test
the robustness and accuracy of the selected putative
signature. This latter strategy is perfectly suited for
the identification of biomarkers of radiation exposure but requires enough samples to be able to split
the data set in two parts. An alternative strategy
consists of leave-one-out cross validation methodologies of the signatures [19-21]. This latter does
not require to split the data-set but could lead to an
overfitting and could destroy the ability of the signatures to generalize beyond the fitting data used.
Description of the available data
Concerning published in vitro studies, we have
identified six publications related to the in vitro
exposure of human lymphocytes or mononuclear
cells. In all of these publications, the expression of
mRNA was measured using cDNA microarrays. It
should be mentioned that the purpose of this review
is not to address publications dealing only with QTPCR but rather to focus on microarray experiments;
therefore, studies performed with the QT-PCR

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technique are not discussed here. With respect to

human blood exposed in vitro, some authors have
exposed whole blood whereas others have chosen to
expose isolated lymphocytes or the CD4+ subset [6].
Moreover, exposure conditions are heterogeneous in
terms of the doses used but also in terms of the time
elapsed between exposure and measurement of gene
expression.
According to the work of Gruel et al. [4] and Mori
et al. [6] it is reasonable to put forward the hypothesis that the different blood subsets do not show the
same response to radiation exposure in terms of
gene modulation. Therefore, the modulations observed in peripheral blood mononuclear cells are an
average of the modulations of different lymphocyte
subsets. In addition to the in vitro exposure of blood
samples, three studies were performed with cells
from skin biopsies: either fibroblasts or keratinocytes [22-24]. In two of those several time points were
analysed [22, 23]. The third study is the only one to
address the impact of dose fractionation [24].
Two papers were found which refer to the chronic
exposure of workers where the physical doses have
been monitored [25, 26]. Gene modulation was
measured on blood samples from these workers for
whom the maximum cumulative doses were lower
than 50 mSv.
Two studies address the modulation of gene expression in skin biopsies obtained from patients
undergoing radiotherapy [27, 28]. Biopsies were obtained from exposed and non exposed skin areas.
Time effect
The knowledge of the post-irradiation evolution
of a biomarker is essential so that dose evaluation
is possible regardless of the delay between exposure
and sampling. An important point regarding the use
of a biodosimeter is that the modifications induced
by ionising radiation do not disappear too fast.
In the majority of in vitro studies the time elapsed
between exposure and analysis varies from 15 min
to 72 h post exposure. Even for this later time, some
significant up regulations are observed [2]. Only one
publication presents data for human lymphocytes
cultured for 7, 17 and 55 days [29]. They found that
the number of genes with an altered expression in
irradiated cells increased with longer culture times.
According to these results it appears possible to to
estimate the dose several days after exposure. Albeit,
the gene modulations were observed following exposure to a rather high dose (3 Gy). Nevertheless, even
with a dose as low as 0.1 Gy, Berglund et al. [27] detected a transient response within the first 24 h after
an in vivo, single radiation exposure of human skin.
Also Franco et al. [22] observed both up and down
regulated genes after 48 and 72 h following exposure
of cultured keratinocytes to 0.01 Gy.
One important notion to keep in mind regarding
the modulation of radiation induced gene expression is that its kinetics varies between different doses

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of radiation in the same biological system. In fact,

in the study of Rodningen et al. [24], the number
of genes modulated either 2h or 24h after in vitro
exposure of skin biopsy to 3.5 Gy is very different.
Only 41 genes were found modulated (28 up regulated probes and 13 down regulated probes) 2 h post
exposure whereas 618 genes were modulated (264 up
regulated probes and 354 down regulated probes) 24
h post exposure. The in vitro study of fibroblasts
also shows that for the 2 Gy dose, the number of
genes regulated 12 h post exposure is nearly 30 times
greater than 6 h post exposure [23]. Interestingly, in
this study a higher number of genes (63 up regulated
genes and 8 down regulated genes) are also modulated 1 h post exposure. We did not find any other
study where modulation of gene expression after
such a short time post exposure was analysed.
Following exposure to low doses of radiation a
significant, transient up-regulation was shown for
zinc finger proteins, keratins, BMP receptor, BAG
and cyclins while a down regulation was detected
for TNF, interleukins, heat shocks proteins and
S100 [27]. These transient responses were obtained
comparing the 3-h and 8-h expression values with
the 0-h and 24-h expression values of mRNA extracted from skin biopsies following in vivo exposure
of patients treated by radiotherapy [27]. In three
other publications dealing with in vitro exposure, an
early transient response following exposure to low
doses of radiation was also observed [2, 15, 23]. One
study was performed with ML1 cells, another with
peripheral blood lymphocytes (PBL) and one with
fibroblasts. In the PBL study, a 4-fold up regulation
of some genes was observed following a dose of 0.5
Gy. This up regulation droped below 2 after a time
of 48 h post exposure. In the study with fibroblasts
an early response was also observed 1 and 3 h post
0.5 Gy exposure. At these time points, the number
of modulated genes was, respectively, 34 and 58,
whereas it dropped to 10 at 6 h post exposure.
Thus, all these studies show that the gene modulation patterns seem closely linked to the time elapsed
between exposure and analysis.
Dose response
Even if a large number of genes is modulated following exposure to ionising radiation, the number of
genes in which the level of expression is proportional
to the dose seems to be low. Indeed the relationship
between radiation dose and modulated expression appeared to be stochastic in most of the genes analysed
by Turtoi et al. [7]. It should be remembered that although microarrays are perfect tools for large scale
screening, an accurate relationship between dose and
the level of expression of various genes can only be
validated with quantitative RT-PCR. Thus, a significant correlation has been found for EGR1; EGR4,
IFN-; c-JUN and TNFSF9 for doses ranging from
1 to 4 Gy. The results presented by Turtoi et al. are
based on six healthy subjects and reflect the inter-in-

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Radiation dose assessment using microarray

dividual variability with clear statistical differences

between the donors. The reason for such differences
has not yet been clarified. In another study, Paul and
Amundson [30] identified the following genes the expression of which increased with the dose 6 h and 24
h post exposure: CDKN1, FDXR, SESN1, BBC3,
PHPT1. In this study, the time post exposure does
not seem to affect the level of the expression of the
genes. In contrast to the results presented by Turtoi et
al, the dose relationship becomes linear starting from
the dose of 2 Gy up to 8 Gy. However, at a lower dose
(0.5 Gy), the relationship is clearly non linear. Non
linearity was also observed by Amundson et al. [2]
4 h post exposure for three genes: DDB2, CDKN1
and XPC. However, with increasing time post exposure the dose relationship approached linearity. For
the longtest time point measured, 72 h post exposure,
an increase in the level of gene expression was still detected but without any clear relationship to the dose.
Several in vivo studies demonstrate that genome
scale measures of gene expression, together with
advanced computational tools, can successfully generate molecular signatures that identify exposure to
clinically relevant levels of radiation in mice and in
humans [19-21]. These molecular signatures consist
of a number of genes specific, for each tested dose
and with a specific level of expression. In general,
there is little or no overlap among the molecular
signatures of each tested dose. The accuracy, specificity and sensitivity of these molecular signatures
are very high but a specific signature seems to be required for each dose.
Sensitivity
The sensitivity is defined as the minimal dose that
can be detected. This depends directly on the intensity of the gene modulation. In the study of Paul
and Admundson [30] the different genes that show a
dose effect relationship do not respond in the same
way. Whatever the dose, FDXR is the gene that is
strongly up-regulated whereas four other genes
(CDKN1, SESN1, BBC3, PHPT1) show a similar
but lower response. The conclusion of this study is
that it is possible to identify individuals exposed to a
dose higher than 0.5 Gy. However several studies described modulations of gene expression in response
to much lower doses [4, 15, 22], even below 0.05 Gy
in haematopoietic cells [4]. These results imply a
regulation of gene expression in response to ionising radiation that depends on the dose and on the
lymphocyte subpopulation. They also demonstrate
the high sensitivity of the regulation mechanism
governing these modulations as measured from 0.05
Gy onwards. Analysis of the overall response suggests that this sensitivity is highest in CD4+ cells, 3
h after exposure. In addition, 70% of the modifications observed 3 h post-exposure to 0.05 Gy were
similar to those measured for the dose 0.5 Gy (72%
for repression and 55% for induction). These similarities in gene expression appear to decrease with

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time. While gene expression tends to recover at 24

h in cells receiving 0.05 Gy, cells exposed to 0.5 Gy
retain a high level of gene modulation.
All these results seem to demonstrate that, in hematopoietic cells, the doses below 0.5 Gy appeared
to produce a short-lived effect which peaks 2 or 3
h after treatment [4, 31]. In addition, it seems that
low doses lead to a more specific cellular response
than high doses. This conclusion is confirmed by in
vivo studies of individuals exposed to radiation due
to the Chernobyl nuclear power plant catastrophe,
where it was possible to detect distinct gene expression patterns in mononuclear cells of individuals exposed to radiation doses in the range of 10 mSv [25].
Another paper describes the use of microarrays in a
group of 14 workers chronically exposed to various
kinds of radiations: X, or -rays [26]. The doses
monitored using physical dosimeters varied from
0.696 to 39.088 mSv with a mean ( SD) of 7.67
10.16. Even with such a low level of cumulated
exposure, the authors identified 78 genes with statistically significant transcriptional changes Among
them 21 were found useful to distinguish exposed
from non-exposed individuals. An unknown parameter in the papers where gene modulation was
measured in cells of chronically exposed individuals
is the time between the end of exposure and blood
sampling. In other words, it is not clear whether the
modulations observed would still be visible several
months/years after the end of exposure.
Conclusions
From this overview of the current literature concerning radiation exposure and microarray approaches, several conclusions and future guidelines
can be highlighted. The major validations of the
gene expression approach for biological dosimetry
are listed in Table 1. It is important to notice that
the exposure conditions found in the different studies are heterogeneous in terms of the doses used
but also in terms of the time between exposure and
analysis. Due to this heterogeneity, it is difficult to
reach consensus with respect to the type(s) of modulated genes.
Nevertheless, it appears that the modulation of
gene expression in response to ionising radiation is
a dynamic mechanism, which depends on multiple
parameters like the dose, the time post-exposure and
the cell type. The studies using microarrays clearly
demonstrate that the expression levels of many
genes are modulated in response to radiation exposure even at very low doses and late post-exposure
times. The current difficulty is that the nature of the
modulated genes seems to evolve with irradiation
conditions. In fact, this literature search was able to
highlight only few genes with reliable correlation between the expression level and the dose. In addition,
these dose-effect correlations change with post-exposure time and seem to have great inter-individual
variations.

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Table 1 | Validation of the characteristics required for a biodosimeter based on the analysis of gene expression
Characteristic

Level of validation

Reference

Effect of gender

No gender effect

[30]

Non invasive

Blood samples: yes

Skin biopsies: no

in vivo: [25, 26]

in vitro: [2, 28]

Dose response

Some individual genes have been tested and present a dose effect relatioship: CDKN1,
FDXR, SESN1, BBC3, PHPT1, EGR1, EGR4, IFN-; c-JUN, TNFSF9, DDB2, XPC

[2, 7, 30]

Identification of dose specific patterns

[23]

Interindividual variations

Yes

[7, 27, 28]

Sensitivity

Around some mSv

[4, 25, 26]

Specificity

Ability to detect chemo vs radio therapies

[21]

Stability

Up to 55 days post exposure (lymphocytes)

Different gene patterns studied for 7 time points up to 72 h (Fibroblast)

[29]
[23]

Chronical exposure

Gene modulation measured for 10 mSv cumulative dose

[25, 26]

Localised exposure

Skin biopsies exposed in vitro show a positive response

Skins samples exposed in vivo show a positive response after the 1 Gy dose

[23, 28]
[27]

Fractionnated exposure

Some common genes between acute and fractionated exposures

[24]

The strength of the genome scale measures of

gene expression is their ability to generate molecular signatures that distinguish clinically relevant
levels of radiation exposure. These molecular signatures consist of dozens of genes specific of each
tested parameter (dose, time, etc.). There is currently little or no overlap between the molecular
signatures obtained for each tested parameter. The
accuracy, specificity and sensitivity of these molecular signatures are very high but a specific signature
seems to be required for each parameter. The ability of a method to recognize a low dose exposure is
particularly important. Moreover, gene expression
modifications induced by low doses will probably
uncover biological phenomena that could potentially be used as a more stable marker of exposure.
The human population is heterogeneous with
respect to health status and medical conditions.
Therefore, it is critical to determine whether the
detected gene modulations following ionising radiation are specific to radiation exposure itself or
whether these signatures are potentially confounded by other stresses. Such level of validation has
been done for mice exposed either to radiation or

to the lipopolysaccharide endotoxin (LPS) and also

for humans exposed to radiation or chemotherapy
drugs [21]. However, these types of studies are still
rare and the confounding influence of many exogenous factors (environmental pollutants for example)
remain to be analysed.
In conclusion, we can expect that the microarray
technology should produce in a near future a reliable
signature of human exposition to low doses of ionising radiation. This signature will probably not be able
to predict a given dose but will rather allow a distinction between exposed and non exposed individuals.
This signature could be helpful in identifying an exposure above a dose threshold, provided that the postexposure time is within a defined period of time. In addition, the described studies open up interesting new
avenues of research for identification of the molecular
mechanisms involved in the dose-specific transcriptional response to ionising radiation. The knowledge
of the molecular mechanisms will perhaps help to
identify persistent fingerprints of radiation exposure.
Submitted on invitation.
Accepted on 16 June 2009.

References
1. Roy L, Roch-Lefevre S, Vaurijoux A, Voisin Pa, Martin
C, Gregoire E, Voisin P. Optimization of cytogenetic
procedures for population triage in case of radiological emergency. Radiat Meas 2007;42:1143-6.
2. Amundson SA, Do KT, Shahab S, Bittner M, Meltzer P,
Trent J, et al. Identification of potential mRNA biomarkers in peripheral blood lymphocytes for human exposure to
ionising radiation. Radiat Res 2000;154:342-6.

ANNALI 3_2009.indb 276

3. Dainiak N, Schreyer SK, Albanese J. The search for

mRNA biomarkers: global quantification of transcriptional and translational responses to ionising radiation.
BJR Suppl 2005;27:114-22.
4. Gruel G, Voisin P, Vaurijoux A, Roch-Lefevre S, Gregoire
E, Maltere P, et al. Broad modulation of gene expression in
CD4+ lymphocyte subpopulations in response to low doses
of ionising radiation. Radiat Res 2008;170:335-44.

8-09-2009 11:30:07

Radiation dose assessment using microarray

5. Kang CM, Park KP, Song JE, Jeoung DI, Cho CK, Kim
TH, et al. Possible biomarkers for ionising radiation exposure in human peripheral blood lymphocytes. Radiat Res
2003;159:312-9.
6. Mori M, Benotmane MA, Tirone I, Hooghe-Peters EL,
Desaintes C. Transcriptional response to ionising radiation in
lymphocyte subsets. Cell Mol Life Sci 2005;62:1489-501.
7. Turtoi A, Brown I, Oskamp D, Schneeweiss FH. Early gene
expression in human lymphocytes after gamma-irradiation-a
genetic pattern with potential for biodosimetry. Int J Radiat
Biol 2008;84:375-87.

radiation exposure in mice and humans. PLoS Med 2007;4:

e106.
20. Gruel G, Lucchesi C, Pawlik A, Frouin V, Alibert O,
Kortulewski T, et al. Novel microarray-based method for
estimating exposure to ionising radiation. Radiat Res 2006;
166:746-56.
21. Meadows SK, Dressman HK, Muramoto GG, Himburg H,
Salter A, Wei Z, et al. Gene expression signatures of radiation response are specific, durable and accurate in mice and
humans. PLoS ONE 2008;3:e1912.

8. Greenstock CL, Trivedi A. Biological and biophysical techniques to assess radiation exposure: a perspective. Prog
Biophys Mol Biol 1994;61:81-130.

22. Franco N, Lamartine J, Frouin V, Le Minter P, Petat C, Leplat

JJ, et al. Low-dose exposure to gamma rays induces specific
gene regulations in normal human keratinocytes. Radiat Res
2005;163:623-35.

9. Zhao W, Chuang EY, Mishra M, Awwad R, Bisht K, Sun L,

et al. Distinct effects of ionising radiation on in vivo murine
kidney and brain normal tissue gene expression. Clin Cancer
Res 2006;12:3823-30.

23. Tachiiri S, Katagiri T, Tsunoda T, Oya N, Hiraoka M,

Nakamura Y. Analysis of gene-expression profiles after
gamma irradiation of normal human fibroblasts. Int J Radiat
Oncol Biol Phys 2006;64:272-9.

10. Mori M, Benotmane MA, Vanhove D, van Hummelen P,

Hooghe-Peters EL, Desaintes C. Effect of ionising radiation
on gene expression in CD4+ T lymphocytes and in Jurkat
cells: unraveling novel pathways in radiation response. Cell
Mol Life Sci 2004;61:1955-64.

24. Rodningen OK, Overgaard J, Alsner J, Hastie T, BorresenDale AL. Microarray analysis of the transcriptional response
to single or multiple doses of ionising radiation in human subcutaneous fibroblasts. Radiother Oncol 2005;77:231-40.

11. Chaudhry MA. Biomarkers for human radiation exposure. J

Biomed Sci 2008;15:557-63.
12. Stears RL, Martinsky T, Schena M. Trends in microarray
analysis. Nat Med 2003;9:140-5.
13. Caldas C, Aparicio SA. The molecular outlook. Nature 2002;
415:484-5.
14. Golub TR, Slonim DK, Tamayo P, Huard C, Gaasenbeek
M, Mesirov JP, et al. Molecular classification of cancer: class
discovery and class prediction by gene expression monitoring. Science 1999;286:531-7.
15. Amundson SA, Bittner M, Meltzer P, Trent J, Fornace AJ,
Jr. Induction of gene expression as a monitor of exposure to
ionising radiation. Radiat Res 2001;156(5 Pt 2):657-61.
16. Amundson SA, Grace MB, McLeland CB, Epperly MW,
Yeager A, Zhan Q, et al. Human in vivo radiation-induced biomarkers: gene expression changes in radiotherapy patients.
Cancer Res 2004;64:6368-71.
17. Amundson SA, Fornace AJ, Jr. Gene expression profiles for monitoring radiation exposure. Radiat Prot Dosimetry 2001; 97:11-6.

25. Albanese J, Martens K, Karanitsa LV, Schreyer SK, Dainiak

N. Multivariate analysis of low-dose radiation-associated
changes in cytokine gene expression profiles using microarray
technology. Exp Hematol 2007;35(4 Suppl.1):47-54.
26. Fachin AL, Mello SS, Sandrin-Garcia P, Junta CM, GhilardiNetto T, Donadi EA, et al. Gene expression profiles in radiation workers occupationally exposed to ionising radiation. J
Radiat Res (Tokyo) 2009;50:61-71.
27. Berglund SR, Rocke DM, Dai J, Schwietert CW, Santana A,
Stern RL, et al. Transient genome-wide transcriptional response to low-dose ionising radiation in vivo in humans. Int J
Radiat Oncol Biol Phys 2008;70:229-34.
28. Goldberg Z, Rocke DM, Schwietert C, Berglund SR, Santana
A, Jones A, et al. Human in vivo dose-response to controlled,
low-dose low linear energy transfer ionising radiation exposure. Clin Cancer Res 2006;12:3723-9.
29. Falt S, Holmberg K, Lambert B, Wennborg A. Long-term global gene expression patterns in irradiated human lymphocytes.
Carcinogenesis 2003;24:1837-45.

18. Ge Y, Dudoit S, Speed TP. Resampling-based multiple testing for

microarray data analysis. TEST 2003;12:1-77.

30. Paul S, Amundson SA. Development of gene expression

signatures for practical radiation biodosimetry. Int J Radiat
Oncol Biol Phys 2008;71:1236-44.

19. Dressman HK, Muramoto GG, Chao NJ, Meadows S, Marshall

D, Ginsburg GS, et al. Gene expression signatures that predict

31. Amundson SA, Do KT, Fornace AJ, Jr. Induction of stress

genes by low doses of gamma rays. Radiat Res 1999;152:225-31.

ANNALI 3_2009.indb 277

277

8-09-2009 11:30:08