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Mutation Research 758 (2013) 29–34

Contents lists available at ScienceDirect

Mutation Research/Genetic Toxicology and
Environmental Mutagenesis
journal homepage: www.elsevier.com/locate/gentox
Community address: www.elsevier.com/locate/mutres

Sulforaphane mitigates genotoxicity induced by radiation and

anticancer drugs in human lymphocytes
Omika Katoch, Arun Kumar, Jawahar S. Adhikari,
Bilikere S. Dwarakanath, Paban K. Agrawala ∗
Division of Radiation Biosciences, Institute of Nuclear Medicine and Allied Sciences, Brig. SK Mazumdar Road, Timarpur, Delhi 110054, India

a r t i c l e i n f o a b s t r a c t

Article history: Sulforaphane, present in cruciferous vegetables such as broccoli, is a dietary anticancer agent. Sul-
Received 28 January 2013 foraphane, added 2 or 20 h following phytohemaglutinin stimulation to cultured peripheral blood
Received in revised form 21 August 2013 lymphocytes of individuals accidentally exposed to mixed  and ␤-radiation, reduced the micronucleus
Accepted 26 August 2013
frequency by up to 70%. Studies with whole blood cultures obtained from healthy volunteers confirmed
Available online 1 September 2013
the ability of sulforaphane to ameliorate -radiation-induced genotoxicity and to reduce micronucleus
induction by other DNA-damaging anticancer agents, such as bleomycin and doxorubicin. This reduction
Keywords:
in genotoxicity in lymphocytes treated at the G0 or G1 stage suggests a role for sulforaphane in mod-
Sulforaphane
Radiation protection ulating DNA repair. Sulforaphane also countered the radiation-induced increase in lymphocyte HDAC
Micronucleus activity, to control levels, when cells were treated 2 h after exposure, and enhanced histone H4 acety-
HDAC inhibitor lation status. Sulforaphane post-irradiation treatment enhanced the CD 34+ Lin− cell population in culture.
Sulforaphane has therapeutic potential for management of the late effects of radiation.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction survival, transformation, and mutation. The accessibility of DNA

Development of radioprotective agents is of interest for appli-
cations in defence, the nuclear power industry, radiation accident
response, space flight, and reducing damage to normal tissues dur-
ing cancer radiotherapy. Progress has been made in development
of prophylactic agents that reduce biological damage when admin-
istered prior to exposure, although amifostine (an amino thiol;
WR-2712, originally developed at Walter Reed Army Institute, USA)
remains the only radioprotective agent approved by the US FDA [1].
Therapeutic management of radiation-exposed persons makes use
of standard supportive care drugs and growth factors, generally
administered following the appearance of symptoms. The devel-
opment of approaches for mitigation [2] of radiation damage has
received attention only recently. An ideal radiomitigating agent
would provide benefit against both acute and delayed effects of
ionizing radiation when administered orally, soon after exposure.
Acute and late effects of ionizing radiation arise due to macro-
molecular damage, especially DNA double strand breaks. Cellular
repair and recovery involve multiple damage response pathways
regulating DNA repair, cell cycle perturbations, cell death, etc., acti-
vated soon after damage induction, and determining subsequent
∗ Corresponding author. Tel.: +91 11 23905187; fax: +91 11 23914390.
E-mail addresses: paban@inmas.drdo.in, pkagrawal@gmail.com (P.K. Agrawala).
in chromatin, an important determinant of response, is regulated
by post-translational modifications (especially acetylation) of
histones, among other factors. Acetylation and deacetylation of
histone and non-histone proteins are tightly regulated by the
opposing effects of HATs (Histone Acetyltransferase) and HDACs
(Histone Deacetylase), respectively [3,4]. Modifiers of HATs and
HDACs affect cellular responses to radiation [4]. HDAC inhibitors
have shown radioprotective activity in animal models, espe-
cially against late effects of radiation [3,5]. Sulforaphane (SFN)
is an HDAC inhibitor present in broccoli [4], SFN also affects the
Nrf-2-Keap system [6,7]. Here, we report the ability of SFN to
mitigate radiation-induced genotoxicity under in vitro conditions,
in cultured peripheral blood samples from a cohort of accidentally
radiation-exposed individuals.
The blood samples used in this study were obtained from
persons referred to INMAS for the assessment of radiation doses
received by individuals who were exposed due to the negligent
dismantling of a Co-60 gamma source in Delhi [8,9]. The gamma
irradiator was imported to India in 1968, with an estimated initial
strength of approximately 3600 Ci, and was disposed in March
2010. The scrap dealer tried to dismantle the instrument, leading
to the exposure of workers and other persons. Some individuals
spent 12–14 h per day near the source, which led to a relatively
higher exposure and subsequent radiation sickness symptoms,
prompting hospitalization of 5–7 persons. The highest estimated

1383-5718/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.mrgentox.2013.08.009
 Author's personal copy
30 O. Katoch et al. / Mutation Research 758 (2013) 29–34
absorbed dose reported for one of the victims was 3.1 Gy. One of
the accidentally exposed individuals succumbed to hematopoietic
syndrome [9].
Our results show that addition of SFN to blood cultures 2 or
20 h post-PHA stimulation (i.e., G0 or G1 stage of the lymphocytes
[10]) significantly decreased micronucleus (MN) formation in the
peripheral blood lymphocytes of these radiation-exposed subjects.
These observations prompted us to investigate further the poten-
tial of SFN as an antimutagen, using blood from healthy volunteers
exposed to agents such as -radiation [11], a known clastogen,
bleomycin [11], a known clastogen, and the radiomimetic agent
doxorubicin [12], a DNA-intercalating agent and topo-II inhibitor.
2. Material and methods
2.1. Chemicals
Sources of materials were as follows: dl-sulforaphane, doxorubicin,
cytochalasin-B, RPMI-1640, IDM, penicillin-streptomycin solution, BCA kit:
Sigma–Aldrich (St. Louis, MO); bleomycin: obtained locally; FBS and PHA: Life
Technologies (India); HDAC assay kit: BioVision, USA (San Francisco, CA); PathScan®
acetyl-histone H4 kit: Cell Signaling Technology, USA (Danvers, MA); PE-Cy-7-
labeled CD-34 antibody: e-Biosciences, USA (San Diego, CA); FITC-labeled lineage
cocktail: BD Biosciences; 7-AAD: Molecular Probes, USA (Eugene, OR). All other
chemicals were purchased locally and were of analytical grade. Sodium-heparin
vacutainers were purchased from Becton Dickinson, USA (San Jose, CA).
2.2. Subjects and sample collection
Blood from healthy volunteers (3–4 ml; n = 6, 4 male and 2 female, ages 25–38
y) and suspected radiation-exposed individuals (n = 32, 26 male and 6 female, ages
12–80) were drawn into Na-heparin-containing vacutainers by trained medical
technologists of the institute. All subjects provided written informed consent. From
questionnaire data, it was found that only ten male individuals (ages 28–56 y) among
the 32 individuals were actually involved in work with the irradiator or spent some
time in its vicinity. The data presented here are from those ten individuals only. Other
donors were relatives who had never visited the site of the irradiator. The guide-
lines provided by the Indian Council for Medical Research (ICMR), the Government
of India, and the institutional ethics committee were strictly followed during this
study, which was duly approved by the Institutional Regulatory Board (IRB Approval
letter INM/TS/IEC/01/1012).
2.3. Whole blood culture
Whole blood for each sample was cultured using a standard procedure. Briefly,
whole blood (0.5 ml) was added to medium (4.5 ml) containing 20% FBS and antibi-
otics, within 2 h of collection of the sample, and stimulated with 100 ␮l PHA (stock
solution 1 mg/ml in PBS). Culture flasks were placed in a humidified CO2 incubator
at 37 ◦ C until harvesting, with intermittent shaking of the flasks every day. Sul-
foraphane (400 nM) was added to each culture either when the lymphocytes were
at G0 (within 2 h of PHA treatment) or G1 (20 h after PHA treatment) phase of the
cell cycle [10].
2.4. In vitro irradiation, genotoxic chemical, and sulforaphane treatment
Blood samples collected from healthy volunteers were cultured as described
above. Cultures were exposed to Co-60 -radiation (0.25, 0.50, 1, or 2 Gy at
1.17 Gy/min) from a Cobalt-60 teletherapy machine (Bhabhatron-II, Panacea Medi-
cal Technologies, Bengaluru, India). Standard chemical dosimetry (Fricke’s method)
and ion-chamber dosimetry were followed for dose calibration. Whole blood
cultures were treated with bleomycin (15 ␮g/ml) or doxorubicin (25 ␮g/ml) in
incomplete medium (without FBS) for 1 h and washed twice with sterile PBS, before
adding complete medium to promote growth and expression of micronuclei. The
treatment conditions were designed to mimic the accidental exposure of indi-
viduals as closely as possible; i.e. test agents were added to whole blood before
mitogenic stimulation. Sulforaphane 400 nM) was added 2 or 20 h after the addi-
tion of PHA, so as to ensure exposure of lymphocytes to sulforaphane at either
the G0 (2 h) or G1 (20 h) stage of the cell cycle [10]. Fig. 1(a–c) depicts schemat-
ically the treatment protocols for each experiment performed. The dose of SFN
was based on our preliminary observation of HUVEC cell survival assessed by the
XTT {2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-
tetrazolium hydroxide} assay (unpublished data).
2.5. Cytochalasin-B-blocked micronucleus assay
The MN assay using cytochalasin-B block was performed essentially according
to the method described earlier [13]. Briefly, all cultures were treated with 5 ␮g/ml
cytochalasin-B 44 h after PHA stimulation until harvesting, i.e., 72 h after PHA stimu-
lation. Cells were washed with PBS twice and fixed with methanol: acetic acid (3:1).
The fixative was removed by centrifugation (300 g; 10 min) and fresh fixative was
added to the cell pellet to remove lysed red blood cells and retain only the nucleated
cells. Fixed cells were kept at 4 ◦ C until slide preparation.
2.6. Slide preparation and scoring
Two or three drops of cell suspension were dropped onto precleaned-chilled
slides and allowed to air dry. Slides were stained with Giemsa diluted in phosphate
buffer, pH 6.8 m for 10–12 min. Excess stain was washed away with the same buffer
and slides were observed under 40× magnification under a light microscope. The
criteria described earlier [13] were followed, for scoring micronuclei and determi-
nation of nuclear division index (NDI). A minimum of 1000 binucleated cells from
three or four slides were scored for each sample and the slides were coded before
scoring.
2.7. Calculation of nuclear division index
The nuclear division index (NDI), a measure of cell division kinetics, was calcu-
lated by scoring cells for the presence of one, two, three or more nuclei. The NDI
was calculated as follows: NDI = (M1 + 2 × M2 + 3 × M3 + 4 × M4)/N, where M1–M4
indicates the number of cells with one to four nuclei, and N is the total number of
cells scored.
2.8. Peripheral blood HSC assay
Blood lymphocytes were isolated using Histopaque and cultured in IDM media
containing 20% FBS and antibiotics, as described earlier [14]. Sets of lymphocyte
cultures (control, 2 Gy, 400 nM SFN, and 2 Gy + 400 nM SFN) were evaluated for
CD34+ Lin− cells between 4 and 48 h. 7-AAD was used to eliminate dead cells from
analysis. A minimum of 50,000 cells from each sample were acquired on a LSR-II
flowcytometer equipped with suitable optics.
2.9. HDAC assay
Blood lymphocytes were isolated using Histopaque and cultured in RPMI media
containing 20% FBS and antibiotics. Nuclear protein samples (10 ␮g) from four sets
of triplicate lymphocyte cultures (control, 2 Gy, 400 nM SFN, and 2 Gy + 400 nM SFN)
were evaluated for deacetylated lysine content using the HDAC colorimetric assay
lit (K331-100; BioVison, San Francisco, CA, USA) at 405 nM, using a Spectra Max 2
multimodal plate reader (Molecular Devices, Eugene, OR, USA), as per the manufac-
turer’s instructions. The values are presented as the amount of deacetylated lysine
equivalent per ␮g nuclear protein, as estimated from a standard curve prepared
using the kit deacetylated lysine standard.
Acetylation status of histone H4 was assayed using PathScan® acetyl histone H4
sandwich ELISA following the manufacturer’s instructions. Cell lysates containing
equal amount of protein from each treatment groups were used after dilution and
absorbance was recorded at 450 nm.
Total protein in the nuclear extract and cell lysate were quantified using the BCA
assay kit (Sigma–Aldrich), with BSA standard.
2.10. Statistical analysis
Data presented are mean ± SD of all samples in the experiment. MN frequencies
were compared using students’ t-test and P < 0.05 was considered significant.
3. Results
3.1. In vitro CBMN assay
A dose response curve for the induction of micronuclei was gen-
erated following in vitro irradiation of human lymphocytes, using
the CBMN assay, in the absorbed dose range 0–2 Gy (Fig. 2). This
curve was used to extrapolate the possible dose received by the
individuals. These individuals did not have a history of smoking,
chronic alcohol consumption, or recent fever. The estimated doses
in nearly ten individuals were in the range 0.25–1.7 Gy, based on
the in vitro dose-response plot (Fig. 2). Addition of SFN (400 nM)
either 2 or 20 h after irradiation resulted in a significant decrease
in MN frequency, which was not strictly dependent on the time of
addition of SFN (Fig. 3). Under the present experimental conditions,
SFN reduced bleomycin- and doxorubicin-induced micronuclei by
30–44% in the whole blood cultures obtained from healthy volun-
teers (Fig. 4). Bleomycin is a radiomimetic agent that can induce
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O. Katoch et al. / Mutation Research 758 (2013) 29–34 31

PHA Fixation
a) 2 or 20 hr 42 or 22 hr 28 hr
Whole blood SFN Cyto-B
Irradiation

Doxo Wash
or and
Bleo PHA Fixation
b) 1 hr 2 or 20 hr 42 or 22 hr 28 hr
Whole SFN Cyto-B
blood

PHA
c)
2 hr 2 hr 20 hr
Lymphocyte SFN
Irradiation Protein Isolation for Protein Isolation for
HDAC assay and H4 H4 Acetylation status
Acetylation status

Fig. 1. A schematic representation of different experimental protocols showing the timings of treatments like (a) whole blood irradiation with respect to PHA stimulation, SFN
treatment and harvesting for fixation, (b) carcinogen (doxorubicin or bleomycin) treatment of whole blood with respect to PHA stimulation, SFN treatment and harvesting
for fixation and (c) isolated lymphocyte irradiation with respect to PHA stimulation, SFN treatment and harvesting for HDAC activity assay. The schematic representation is
not to scale and details are given in the text.

clustered DNA damage; doxorubicin is a topo-II poison capable that
can cause DNA strand breaks.
Treatment with 400 nM SFN, either 2 or 20 h after PHA stim-
ulation, significantly (P < 0.05) reduced the radiation-induced MN
frequency in the lymphocytes of the ten subjects who worked in
the contaminated area (Table 1). Similar results were obtained
with in vitro irradiation of the blood from normal volunteers
(Fig. 3).
The concentration of SFN (400 nM) used here did not compro-
mise the nuclear division index (NDI), either in case of the in vitro
irradiation of whole blood from healthy volunteers or the blood
cultures from suspected exposed individuals (Table 1).
3.2. CD 34+ Lin− cell population in peripheral blood
A significant decrease in the population of CD 34+ Lin− cells was
observed in the group receiving 2 Gy irradiation, at all time points
studied, as compared to the control group (Fig. 5). The SFN and
irradiation plus SFN groups were comparable to the control group
at corresponding time points.
3.3. HDAC activity assay
Irradiation (2 Gy -radiation) of the lymphocytes significantly
(P < 0.05) increased the amount of deacetylated lysine in the nuclear

250
2
R = 0.98
200
MN per 1000 BN

150

100

50

0
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2
Radiation Dose (Gy)

Fig. 2. Heparinized blood from healthy six volunteers was cultured and exposed to 0–2 Gy Co-60 gamma rays before PHA stimulation. Slides were prepared for MN observation
as described in the text. MN frequencies in all treatment groups were plotted as a function of radiation dose. The curve obtained without SFN treatment was used as the
standard curve for absorbed dose estimation in the accidentally exposed individuals, the dotted lines shows upper and lower confidence limits (95% confidence interval).
 Author's personal copy

32 O. Katoch et al. / Mutation Research 758 (2013) 29–34

Fig. 3. Heparinized blood from healthy volunteers was cultured and exposed to 0–2 Gy Co-60 gamma rays before PHA stimulation. Whole blood cultures exposed to identical
radiation dose were treated with 400 nM SFN either 2 or 20 h post-PHA stimulation. 72 h after culture, slides were prepared for MN observation as described and MN
frequencies in all treatment groups were plotted as a function of radiation dose. The curve obtained without SFN treatment was used for absorbed dose estimation in the
accidentally exposed individuals. Values presented are mean ± SD of blood from six healthy volunteers.

Table 1
Effect of SFN treatment of radiation induced MN frequency and NDI.

MN frequency (Percent) NDI

No SFN SFN Fold change No SFN SFN Fold change

1.30 1.20 0.92 1.56 1.50 1.04

1.69 1.20 0.71 1.55 1.56 0.99
1.40 1.15 0.82 1.56 1.56 1.00
1.20 1.11 0.92 1.45 1.50 1.03
1.40 1.25 0.89 1.48 1.51 1.02
0.90 0.90 1.00 1.56 1.52 0.97
5.15 2.80 0.54 1.58 1.44 1.09
4.45 3.57 0.80 1.60 1.67 0.96
4.10 3.14 0.77 1.55 1.49 1.04
4.65 3.60 0.77 1.56 1.48 1.05
24.90 7.20 0.29 1.46 1.53 0.95
17.30 10.00 0.57 1.44 1.45 1.006
8.70 7.20 0.29 1.46 1.53 0.95
Fig. 4. Whole blood samples from healthy volunteers were treated with chemical 5.50 3.10 0.56 1.55 1.52 0.98
mutagens like bleomycin or doxorubicin for 1 h, washed with sterile PBS and treated 2.90 2.20 0.75 1.48 1.49 1.006
with PHA. SFN was administered at a concentration of 400 nM at either 2 or 20 h post 4.50 3.25 0.72 1.51 1.50 1.006
PHA stimulation and MN frequency was estimated as described in the text. Values Whole blood cultures from accidentally radiation exposed individuals were treated
presented are mean ± SD of blood from six healthy volunteers. with 400 nM SFN 2 h after PHA stimulation. First six samples are from healthy vol-
unteers and remaining are from the cohort of accidentally exposed individuals.
Micronuclei frequency in binucleated lymphocytes and nuclear division index were
estimated as described in Section 2. Effect of SFN was designated in terms of fold
change (ratio of MN frequency or NDI with out SFN and with SFN treatment groups).

extract as compared to the untreated samples (controls), at 2 h

post-SFN treatment (Fig. 6a). SFN alone did not alter the total
deacetylated lysine level while treatment with 400 nM SFN 2 h after
2 Gy irradiation restored the deacetylated lysine level comparable
to the untreated controls (Fig. 6a).
Fig. 6b depicts the changes in acetylation status of histone H4
as a function of time. SFN treatment 2 h after 2 Gy irradiation was
observed to enhance H4 histone acetylation status until 24 h, as
compared to the control and radiation-alone groups.

4. Discussion

Fig. 5. The percentage of live circulating hematopoietic stem cells were studied The mutagenic effects of physical and/or chemical agents can be
using antibodies for CD 34 and Lin markers and 7-AAD as described in Section 2. ameliorated by multiple mechanisms, broadly classified as mod-
Values presented are mean ± SD of blood from six healthy volunteers. ification (prevention) of the induction of genomic damage and
enhancement of DNA repair capability. Several compounds can

a 30
25
20
HDAC Acvity 15
10
0
b 4
3.5
3
Acetylated H4 (O.D)
2.5
1.5
0.5
0
1hr
Fig. 6. (a): HDAC activity (␮M deacetylated lysine generated per ␮g nuclear protein) in the lymphocyte nuclear protein obtained from various treatment groups as depicted
in the figure was assayed as described in Section 2. Values presented are mean ± SD of blood from six healthy volunteers. (b) The acetylation status of histone H4 was studied
using PathScan acetyl H4 sandwich ELISA as described by the manufacturer. Values presented are mean ± SD of blood from six healthy volunteers.
antagonize the effects of mutagenic agents, and these are grouped
based on their mechanism and site of action. For example, inhibitors
of tumor initiation are called blocking agents of mutagenesis,
while suppressing agents are inhibitors of cancer promotion and
progression [15]. On the other hand, desmutagens inactivate muta-
gens before they can attack DNA and antimutagens interfere with
the fixation of DNA damage [16]. The ICPEMC Expert Group on
Antimutagens and Desmutagens made a distinction between stage-
1 inhibitors, acting extracellularly, and stage-2 inhibitors, acting
intracellularly [17]. Antimutagens act at the cellular level as mod-
ulators of DNA repair and replication [18]. The protective effects of
antimutagens can be mediated by increases in the fidelity of DNA
replication, by stimulating error-free repair of DNA damage, or by
inhibiting error-prone repair systems.
As was evident from the in vitro irradiation studies, SFN ame-
liorated radiation-induced MN formation under various treatment
regimens, implying its potential application as an antimutagen. MN
formation is a result of DNA strand breaks caused by radiation and
other clastogenic agents. Amelioration of MN formation by SFN
administered 2 or 20 h following PHA stimulation (Figs. 3 and 4)
suggests that its anti-mutagenic effect could arise either due to
enhanced repair or reduced manifestation of the damage. Reduc-
tion in the micronuclei by SFN was not due to reduced proliferation
(an important pre-requisite for MN expression), as revealed by the
Author's personal copy
O. Katoch et al. / Mutation Research 758 (2013) 29–34 33
Con 2Gy SFN DR
Treatme nt groups
2hr 4hr 24 hr 1hr 2hr 4hr 24 hr 4hr 24 hr 4hr 24hr
Con 2Gy SFN 2Gy+SFN
Treatments
ND index values (Table 1). Another study [19] also demonstrated
the antimutagenic potential of SFN against four known mutagens,
without any adverse affects of SFN on cell division or cell death at
a concentration of >10 ␮M. In our study also, SFN did not induce
significant apoptotic cell death (data not shown), which otherwise
could contribute to the reduction in micronuclei frequency. These
observations also suggest that the SFN concentration required for
mitigation (as used here) is neither toxic to cells nor has any cyto-
static effect.
Ex vivo SFN treatment many days to several weeks after
accidental exposure to radiation was effective in mitigating
radiation-induced micronuclei in the blood cells of human sub-
jects (Table 1). It is important to note here that the individuals
were exposed to low doses of gamma and beta (since the pencils
were dismantled) radiation, over a period of several days inter-
mittently before sample collection took place. The total absorbed
dose and dose rate were dependent on the time spent and prox-
imity of the individuals to the source, based on the nature of their
work duties. This indicates that SFN can ameliorate micronuclei
arising not only due to exposure to acute gamma radiation but also
due to mixed radiation exposure over a prolonged period. If SFN
merely suppressed the expression of micronuclei and not reduce
the manifestation of damage, then the survival of cells is expected
to be compromised under these conditions, which should affect
 Author's personal copy
34 O. Katoch et al. / Mutation Research 758 (2013) 29–34
mortality/morbidity of the exposed individual. Taken together,
these observations suggest that SFN indeed mitigates radiation-
induced damage by modifying genomic damage in cells. SFN has
been reported to act via the Nfr-2 keap1 pathway to impart its anti-
cancer properties [20]. More recently, SFN has been observed to act
as a HDAC inhibitor [21], which can alter the acetylation status of
histones and non-histone proteins, imparting significant effects not
only on chromatin structure but also on signal transduction path-
ways. The current study has shown the ability of SFN to restore
normal levels of deacetylated lysine in the nuclear proteins (HDAC
activity), which was increased due to radiation exposure (Fig. 6a).
This in turn suggests a reduced amount of acetylated histones
in the irradiated samples and an increased amount of acetylated
histones in the irradiated-plus-SFN-treated group. Increased acet-
ylation in the later may facilitate the downstream events like
transcription and repair of the more open DNA [22], which ulti-
mately may result in lesser cytogenetic damage. Reduction in the
MN frequency observed in the peripheral blood lymphocytes of
individuals who earlier (several weeks before actual blood sample
collection) were exposed to radiation accidentally suggests that the
residual genomic damage is susceptible to alterations or manipu-
lation by HDAC linked changes in chromatin status. Our further
observation on the prolonged hyperacetylation status of histone
H4 (Fig. 6b) also indicates an increased repair activity following
SFN treatment. H4 acetylation status facilitates DNA double-strand
break repair [23,24]. Whether the anticancer properties of SFN are
linked to its HDAC inhibitory effects needs further investigation.
Irrespective of the mechanisms underlying the reduction in cyto-
genetic damage, our observations suggest that SFN can reduce both
acute as well as late effects related to radiation-induced genomic
damage.
5. Funding
The work was supported by the Defence Research and Develop-
ment Organization, Government of India.
Conflicts of interest
No conflict of interest exists.
Acknowledgments
We thank V.R. Senthil and Namita Kalra for carrying out the dose
calibration of the irradiator and for helping with flow cytometry.
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