Mutation Research 764–765 (2014) 46–57

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

MicroRNAs as potential biomarkers in diseases and toxicology

Bénazir Siddeek a,b,f , Lilia Inoubli a,b , Nadjem Lakhdari a,b , Paul Bellon Rachel a,b ,
Karma Claire Fussell g , Steffen Schneider g , Claire Mauduit a,b,c,d ,
Mohamed Benahmed a,b,e,∗
a
Inserm, U1065, Centre Méditerranéen de Médecine Moléculaire (C3M), Team 5, Nice, F-06204, France
b
Université de Nice Sophia-Antipolis, UFR Médecine, Nice, F-06000, France
c
Université Lyon 1, UFR Médecine Lyon Sud, Lyon, F-69921, France
d
Hospices Civils de Lyon, Hôpital Lyon Sud, laboratoire d’anatomie et de cytologie pathologiques, Pierre-Bénite, F-69495, France
e
Centre Hospitalier Universitaire de Nice, Pôle Digestif, Gynécologie, Obstetrique, Centre de Reproduction, Nice, F-06202, France
f
BASF Agro, Ecully F-69130, France
g
BASF SE, experimental toxicology and ecology, 67056 Ludwigshafen, Germany

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

Article history: MiRNAs (microRNAs) are single-stranded non-coding RNAs of approximately 21–23 nucleotides in length
Received 7 January 2014 whose main function is to inhibit gene expression by interfering with mRNA processes. MicroRNAs
Received in revised form 20 January 2014 suppress gene expression by affecting mRNA (messenger RNAs) stability, targeting the mRNA for degra-
Accepted 20 January 2014
dation, or both. In this review, we have examined how microRNA expression could be altered following
Available online 30 January 2014
exposure to chemicals and how they could represent appropriate tissue and more interestingly circu-
lating biomarkers. Among the key questions before using the microRNA for evaluation of risk toxicity, it
Keywords:
remains still to clarify how they could be causally involved in the adverse effects and how stable their
MicroRNAs
Chemicals
changes are.
Circulating biomarkers © 2014 Published by Elsevier B.V.

1. Introduction In animals, siRNA typically binds perfectly to its mRNA target, a

Small RNAs of 20–30 nucleotides continue to be discovered,
including some that are specific to plants or animal lineages (for
reviews, see [1,2]). Various small RNAs with distinctive charac-
teristics have been identified and can be classified into three
classes based on their biogenesis mechanisms, the type of RNA
protein that they are associated with and the nature of their tar-
gets: microRNAs (miRNAs), endogenous small interfering RNAs
(endo-siRNAs) and Piwi-interacting RNAs (piRNAs). Endogenous
siRNA are short double-stranded RNAs that contains 21–23 nucleic
acids with a 19-nucleotide duplex region which is able to inhibit
the gene expression of specific proteins by a mechanism called
RNA interference. The piRNAs are highly enriched in the animal
germline only and originate from single-stranded precursor RNAs
[3]. MiRNAs are single-stranded non-coding RNAs of approximately
21–23 nucleotides in length which main function is to inhibit gene
expression by interfering with messenger RNA (mRNA) processes.
∗ Corresponding author at: INSERM (National Institut of Health and Medical
Research), U1065, Team 5, Bâtiment Universitaire Archimed, Centre Méditerranéen
de Médecine Moléculaire (C3M), 151 route Saint-Antoine Ginestière, BP 2 3194,
06204 Nice Cedex 3, France. Tel.: +33 492 035 630; fax: +33 492 035 640.
E-mail address: benahmed.m@chu-nice.fr (M. Benahmed).
perfect match to the sequence, whereas miRNA can inhibit the
translation of many different mRNA sequences because its pair-
ing is imperfect. MiRNAs suppress gene expression by affecting
mRNA stability, targeting the mRNA for degradation, or both [4].
Near to 1872 miRNAs have been identified in human with the
potential to regulate the expression of about two-third of human
mRNAs and influence almost all genetic pathways. The possibilities
of predictions for miRNA-target recognition have been developed
using computer modeling [5]. A growing number of data show that
microRNAs are involved in the expression of genes involved in
xenobiotic exposure [6–8]. Better understanding of the non-coding
RNAs is critical to determine its potential key role in the impact of
environmental factors and particularly xenobiotic exposure [9] (for
a review, see [10]).
Since their discovery, there is a growing interest in miRNAs
research allowed by the development of efficient and sensitive tools
for their study. MiRNAs can be extracted from tissues and quanti-
fied easily by RT-qPCR even with low amount of input material [11].
They can also be detected by hybridization (Northern, Panomics,
Nanostring, MicroArrays) ([12,13] or by Next generation sequenc-
ing [14]. Since a decade, a number of studies are pointing out the
role of miRNAs in the regulation of key cellular processes and the
appearance of pathologies. MicroRNAs (miRNAs) have been impli-
cated in posttranscriptional regulation of many gene expressions

1383-5718/$ – see front matter © 2014 Published by Elsevier B.V.

http://dx.doi.org/10.1016/j.mrgentox.2014.01.010
 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57 47

Table 1
Effects of drugs, natural and chemical compounds on miRNAs expression.

Compound Organ/Cell Species Altered miRNAs References

Cigarette smoke Spermatozoa Human Mir-340, mir-365, 129-3p, 634 [68]

Volatile organic compounds Lung Mouse miR-1187, miR-125a-3p, miR-125b-5p, miR-466c-5p, [100]
(formaldehyde, benzene, toluene, and miR-5105, miR-3472
xylene)
Trichostatin A Primary hepatocyte Rat miR-379, miR-143, miR-122, miR-122, miR-143, miR-379 [101]
Sodium arsenite TK6 cell line miR-222, miR-210, [102]
Arsenic trioxide T24 cell line mir-19a [103]
Cadmium Human Mir-146 [95]
Aluminum-sulfate HN cells Mir-146 [104]
Aluminum-sulfate HN cells miR-9, -128, -125b [105]
Air pollution metal-rich PM Leucocytes Human Mir-222, mir-21 [106]
Cigarette smoke Lung Rat Mir-294, let-7c, miR-34c, -222 [107]
Cigarette smoke Lung Human Mir-218 [108]
4-(methylnitrosamino)-1-(3-pyridyl)-1- Lung Rat Mir-126, Mir-34 [109]
butanone (NNK) (tobacco
carcinogen)
Hexahydro-1,3,5-trinitro-1,3,5-triazine Liver Mouse let-7, miR-15, -16, -26, -181, miR-10b [110]
Hexahydro-1,3,5-trinitro-1,3,5-triazine Neuron Mouse miR-206, -30, -195 [110]
Carbon tetrachloride Hepatocytes Rat miR-298, -370 [111]
Dioxins Hepatocytes Rat Mir-191 [112]
Bisphenol A 3A placental cell Human Mir-146a [113]
Wy-14, 643 Hepatocyte Mouse Let-7c [114]
Estradiol benzoate Testis Rat Mir-29 [66]
Ethanol Neurons Mouse Mir-21, mir-335,mir-153 [115]
Tamoxyfen Hepatocyte Rat Mir-17-92 cluster, mir-106a, mir-34 [116]
Acetaminophen; carbon tetrachloride Hepatocytes Mir-298, mir-370 [111]
Iron – and aluminum – sulfate Neuron Human Mir-9, mir-125b, mir-128 [105]
5-fluorouracil (5-FU) (drug) Colon Human Mir-200b [117]
Residual oil fly ash Heart Rat miR-1 and miR-133, miR-21, miR-24, and miR-29 [118]
Vorinostat, myo-inositol, bexarotene, Lung Mouse [119]
pioglitazone (cigarette smoke)
6-Mercaptopurine (immunosuppressive Placenta Rat miR-195, miR-21, miR-29c and miR-34a, 146bmiR-144 [120]
drug) and miR-451
4-(methylnitrosamino)-1-(3-pyridyl)-1- Lung Rat miR-206 and miR-133b [121]
butanone
Radon Lung BEAS2B cell line Human hsa-miR-483-3p, hsa-miR-494, hsa-miR-2115*, [122]
hsa-miR-33b, hsa-miR-1246, hsa-miR-3202, hsa-miR-18a,
hsa-miR-125b, hsa-miR-17*, and hsa-miR-886-3p
7,12-dimethylbenz(␣)anthracene (DMBA) In the liver, spleen and Mouse miR-34a and miR-155 miR-21 [123]
and N-methyl-N-nitrosourea (MNU) kidneys
Acetaminophen, allyl alcohol, and Liver Rat miR-122 [124]
␣-naphthyl isothiocyanate
2,3,7,8-tetrachlorodibenzo-p-dioxin Thymus Mouse miR-122 and miR-181a miR-23a, miR-18b, miR-31 [125]
(environmental contaminant) miR-182
2,3,7,8-tetrachlorodibenzo-p-dioxin Embryo Zebrafish miR-27e [126]
Doxorubicin Heart Mouse 208b, miR-216b, miR-215, miR-34c and miR-367 [127]
Cyanobacterial hepatotoxin microcystin-LR Whitefish let-7c, miR-9b), (miR-16a, miR-21a, miR-34a) (miR-122) [128]
Ethanol Liver Rat Mir-21 [129]
Printex 90 carbon black nanoparticles Lung Mouse miR-135b [130]
Arsenic Umbilical endothelial Human [128]
cell
Arsenic Liver Mouse [131]
Acetaminophen Liver Mouse 74-5p, 135a*, 466g, 1196, 466f-3p, 877, 342-3p, 195, 375, [132]
29c, 148a, 652
Gentamicin Kidney Mouse Mir-21, 155 [133]
2-Amino-1-methyl-6-phenylimidazo[4,5- Colon Rat Let-7 [134]
b]pyridine (carcinogen from cooked
meat)
Perfluorooctane sulfonic acid Brain Rat miR-466b, -672, and -297 [135]
Cadmium HepG2, liver Human Let-7 [136]
Cisplatin Hela [137]
Folate deprivation and arsenic exposure Lymphocyte Human Mir-222 [102]
Benzo[k]fluoranthene Hepatocyte Human miR-146a, miR-365, let-7f, miR-199b-5p,miR-30c-1* [138]
Thapsigargin, deoxycholic acid Hepatocytes Mouse mir-199a-5p [139]
Fenhexamid and fludioxonil (fungicide) Breast cancer cells Human Mir-21, miR-125b and miR-181a [140]
DDT and BPA Breast cancer cells Human Mir-21 [141]
Nonylphenol Sertoli cells, testis Mouse miR-135a* and miR-199a-5p [142]
Pegylated leptin antagonist Hypothalamus Rat miR-10a, miR-200a, rno-miR-409-5p, miR-125a-3p [143]
Lycopene Liver Rat Mir-21 [144]
Licorice flavonoid Liver Mouse Mir-122 [145]
Quercetin Liver Mouse miR-122 and miR-125b [146]
Ethanol Liver Mouse Mir-217 [147]
Allyl-isothiocyanate Macrophage Mouse Mir-155 [148]
High fat diet Adipocytes Mouse Mir-21 [149]
Conjugated linoleic acid Adipocytes miR-103 mir-107, miR-221, mir-222 [150]
High fat diet Adipocytes Mouse Mir-143 [151]
48 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57

and control of different processes such as apoptosis, DNA repair,

oxidative stress response, cancer and cellular development. Those
discoveries in miRNAs research open new insights in toxicology and
raise the question of miRNAs regulation in response to exposure to
environmental chemical toxicants or natural compounds such as
alcohol, smoke, stress, hormones, medication, and diets (Table 1).
This would facilitate the bio-monitoring and preventive strategies
for drug development, and clinical pharmacotherapy.
This review describes the current knowledge on the miRNAs
expression, their potential changes in toxicological studies and
circulating miRNA as potential toxicological biomarkers will be
discussed.

2. MiRNA expression (Fig. 1)

The stem-loop structures from which miRNAs are derived

are disseminated throughout the genome, either within intronic
sequences of protein-coding genes, within intronic or exonic
regions of non-coding RNAs, or set between independent tran-
scription units (inter-genic). The location of some of these intronic
miRNAs is evolutionarily conserved and they are similarly coex-
pressed with their host genes in different animals. Here, we review
the recent advances in miRNA biogenesis in animals. Particularly,
we focus on the roles of miRNAs in vertebrate physiology. The
majority of intronic miRNAs are transcribed from the same pro-
moter as the host gene. However, approximately 35% of intronic
miRNAs have upstream regulatory elements consistent with pro-
moter function [15]. Pri-miRNAs are mostly transcribed by RNA
polymerase II (Pol II) [16], and to a less extent by Pol III [17].
Their promoters also contain transcription start sites, CpG islands,
expression sequence tags, and conserved transcription factor bind-
ing sites, enhancers and silencers [18,19]. MiRNA biogenesis is
based on series of processing steps to convert the primary miRNA
(pri-miRNA) transcript into the biologically active, mature miRNA
(Fig. 1), [for a review see [1,20]]. Firstly, the transcribed pri-miRNA
is cleaved by the RNase III-like enzyme Drosha in the nucleus [21]
generating a miRNA precursor about 60–70 nt called pre-miRNA.
Cleavage by Drosha requires the co-factor DGCR8 (DiGeorge critical
region 8), also known as Pasha [22]. The miRNA biogenesis machin-
ery is also subjected to regulation by feedback loops, as evidenced
for the Drosha–DGCR8 complex [23,24]. Once produced, the pre-
miRNA is translocated to the cytoplasm through the nuclear pore
complex by Exportin-5. In the cytoplasm, the pre-miRNA hairpin
associates with the RNase III-like enzyme Dicer that cleaves it into
a double-stranded miRNA duplex comprised of the mature miRNA
and the miRNA* (also known as passenger strand) [25,26]. The
guide strand functions as a mature miRNA and is incorporated into
an RNA-induced silencing complex (RISC). This complex contains
an Argonaute (Ago) protein as a primary component that binds to
the target mRNA and degrades the passenger strand. Mature miRNA
guides RISC to recognize target sequences located in the 3 UTR of
mRNAs leading to the inhibition of translation or degradation of
mRNA [27]. MiRNA biogenesis can be regulated at each step by
various mechanisms. At the transcriptional level, a number of fac-
tors like c-myc, or HMGA1 have been showed to regulate miRNAs
transcription, and have been reported in databases such as Trans-
mir [28]. Further steps, like the nuclear processing of miRNAs by
Drosha can be also modulated by the transforming Growth Factor
␤ (TGF␤) signaling pathway, the Smads [29]. The c-Myc oncogenic
transcription factor also transactivates drosha mRNA expression
thus upregulating the Drosha protein level in addition to the tran-
scriptional regulatation of miRNAs [30]. Tumor suppressor p53
promotes some pri-miRNA processing via interaction with p68 [31].
Furthermore, it has been shown that the p38 MAPK-MK2 signaling
pathway promotes miRNA biogenesis by facilitating the nuclear
Fig. 1. MiRNA biogenesis. MicroRNA (miRNA) genes are transcribed by RNA poly-
merase II (Pol II) to generate the primary transcripts (pri-miRNAs). Cleavage of the
pri-miRNA occurs within the nucleus by Drosha and DGCR8 complex (also known as
the Microprocessor complex) which interact with helicases p68 and p72, to generate
a ∼65 nucleotide pre-miRNAs. Pre-miRNA has a ∼2-nt 3 overhang, which is recog-
nized by the nuclear export factor exportin 5. Once export through the nuclear pore
complex into the cytoplasm, the RNase III Dicer supported by the proteins TRBP or
PACT catalyses the second processing step to produce miRNA duplexes. Dicer, TRBP
or PACT and Argonaute (AGO 1–4) mediate the processing of pre-miRNA and the
assembly of the RISC (RNA-induced silencing complex). One strand of the duplex is
preferentially retained by the Ago protein as the mature miRNA, whereas the other
strand is degraded. The complex contains an Ago protein, GW182 and mature miRNA
which is required for gene silencing, which act by specifically binding to the 3 -UTR
regions of target mRNAs, causing translational repression or mRNA degradation.
localization of p68 [32]. The nuclear-cytoplasmic exportation can
be regulated by steroids like estrogen and progesterone which
increase Exportin-5 mRNA expression [31]. The tumor suppres-
sor, BRCA1 (breast cancer susceptibility gene 1), associates with
Microprocessor complex to accelerate processing of pri-miRNAs
specifically associated with cancer [33]. Other proteins such as
Esr1 (also known as estrogen receptor alpha) [34], NF90 and NF45
(nuclear factor 90 and 45) [35] associate with Drosha and inhibit
also the processing of specific pri-miRNAs such as let-7 family
members.
Proteins regulating pri-miRNA processing by recognition of the
stem-loop sequence or structure like hnRNP-A1 (heterogeneous
nuclear ribonucleoprotein A1), KSRP (KH-type splicing regulatory
protein) have been also been described. Lin28 (abnormal cell lin-
eage factor 28) is an interesting protein involved in regulating
multiple aspects of miRNA biogenesis. This protein can inhibit pri-
let-7 processing [36,37] by sequestering pri-let-7 miRNAs in the
nucleoli away from the Microprocessor. Interestingly, hnRNP-A1
protein can also interact with pri-let-7a, but in this case, it nega-
tively regulates the processing [38].
 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57
The terminal loop region is a determinant of microRNA biogene-
sis. This region is crucial for both positive (for example, hnRNP A1)
and negative (for example, Lin28) regulators to modulate miRNA
levels and thereby gene regulation, reviewed in [39]. Finally, miRNA
stability and degradation are dependent on different mechanisms
that give to the majority of mature miRNAs, a half-life in the order of
several hours to several days [40,41]. This suggests that a cis-acting
element in the sequence of mature miRNA regulates specificity
of degradation of miRNAs. Studies in several model systems have
identified the molecular mechanisms of miRNA-regulated decay.
The animal miRNAs generally lack a protective 2 -O methyl group
at their 3 terminus and display template-independent nucleotide
addition, mostly adenylation or uridylation that may regulate
miRNA stability [42].
The XRN (5 –3 exoribonuclease) family of enzymes and
hPNPaseold-35 (human polynucleotide phosphorylase protein)
play various roles in miRNA stability.
In the context of the present brief review aiming at identifying
the role of miRNA expression in toxicology, it will be of major inter-
est to in the future to identify how drugs/chemicals may affect this
miRNA biosynthesis and degradation pathways.
3. MiRNA in toxicology (Table 1)
Recently, the identification of miRNAs targets highlighted the
roles of miRNA in the posttranscriptional regulation of cytochrome
P450 (CYP) and nuclear receptors and considered their poten-
tial relevance and application for toxicological studies. P450s are
important enzymes that catalyze the metabolism of xenobiotics
including drugs, environmental chemicals, and carcinogens. The
transcriptional regulation of CYPs by nuclear receptors has been
well studied and the central role of miRNAs in the regulation of
CYPs and nuclear receptors has been evidenced [43]. For example,
CYP1A1/1A2 is regulated by miR-142-3p and miR-200a, CYP2C19
by miR-34a, CYP2D6 by let-7b and CYP2E1 by miR-10a and let-
7g [44]. In addition, miRNAs can modulate the action of nuclear
receptors. Nuclear receptors are ligand-activated transcription fac-
tors that regulate the expression of target genes by binding to
their promoters. The expression of cytochrome P450 is regulated
by receptors such as pregnane X receptor (PXR), the aryl hydro-
carbon receptor (AhR), and the constitutive androstane receptor
(CAR). Those receptors are targeted by a number of miRNA and
represent a way of P450 regulation. Indeed, mir-148a, which is
abundantly expressed in the liver, targets the human PXR [45].
It was reported that miR-375 with complex regulatory effects on
AhR mRNA could mediate in part the action of pollutants such
as ambient particulate matter and diesel exhaust particles in the
induction of asthma [45]. Also, the activation of AhR by Tranilast,
an Anti-allergy Drug was shown to facilitate cell reprogramming
in a miR-302-dependent way [46]. Shizu et al. suggested that
phenobarbital-mediated down-regulation of miR-122 is an early
and important event in the AMPK-dependent CAR activation and
transactivation of its target genes [47]. Thus, microRNAs appear
as key players in the regulation of genes involved in absorption,
distribution, metabolism, and excretion.
Peroxisome-proliferator-activated receptors (PPARs) are
ligand-activated nuclear receptors that exert in the liver a tran-
scriptional activity. PPAR␣ was reported to be specifically regulated
by miR-21 and miR-27b, in the liver [48]. Additionally, miR-10b
was shown to target PPAR␣ and induce steatosis in hepatocytes
[49]. Indirectly, PPAR␥ activity appears to be under the control
of miR-132. Inversely, miRNA expression profiling indicated that
activated PPAR␣ is a major regulator of hepatic miRNA expres-
sion and revealed that let-7C signaling cascade is critical for
PPAR␣ agonist-induced liver proliferation and tumorigenesis [50].
49
Concerning AhR, its activation in TH17 cells regulates miR-132/212
cluster and induces the cells differentiation [51]. Among nuclear
receptors that are regulated by miRNAs, Esr1 was reported as a
target of miR-206 [52], miR-221, miR-222 [53], and miR-22 [54].
Conversely, miRNAs can also be regulated by nuclear receptors
at the transcriptional level and during their maturation [55].
Some authors pointed out the potential role of these miRNAs in
anti-estrogen therapies. In the mouse testis, miR-184 was shown
to target nuclear receptor corepressor 2 (Ncor2) and by this way,
regulates mammalian spermatogenesis [56].
Considering the wide actions of miRNAs on the expression
of genes in key processes involved in detoxification, chemical
metabolic activation, and in the activity nuclear receptor, miRNAs
studies open new insights in the mode of action of drugs, hormones
or endocrine disrupting chemicals.
In this way of elucidating miRNAs function, the understand-
ing of cell-type specificity of miRNA expression is an important
step. However, very few miRNAs are exclusively found in indi-
vidual tissues or cells. The miRNA expression varies from highly
specific to ubiquitous and, for conserved miRNAs, is comparable
between rodents and human [57]. One example of tissue specificity
was found in the lung where miR-195 and miR-200c were specif-
ically expressed in the rat lung [58]. In few research areas such as
hepato-toxicity or cardiology, specific miRNAs have been identified
as markers of specific tissue injury. Indeed, miR-208 is described as
a myomiR since it is found at high levels in cardiac tissue. MiR-208
expression is deregulated in various cardiovascular diseases. More-
over, its expression in the cardiac tissue is described as a sensitive
biomarker for cardiac injury in human [59], and inhibition studies
concluded that it might be an interesting therapeutic target [60].
More often, this is a miRNAs expression profile that is linked to a
specific pathology. For instance, Shapiro et al. have recently exam-
ined the role of miRNAs in renal Ischemia reperfusion injury using
expression profiling. They found that samples that underwent renal
injury can be distinguished from controls based on the alterations
in nine miRNAs [61].
Interestingly, assessment of Doxorubicin-induced cardiomy-
opathy versus anthracyclins in rats indicated that miRNAs
expression profile could give indications about the specific tissue
response in a dose and a time dependant manner [62]. In this tissue,
they represent early toxicity indicators, since their modifications
are detected earlier than changes in gene expression. Thus, miRNAs
can help to better understand chronic/acute toxicity mechanism,
since they may be regulated earlier than genomic toxicological
markers. Similarly, MCF7 breast cancer cells showed a dose- and
time-dependent modification in the miRNA expression levels after
a treatment with the chemotherapeutic drug 5-flourouracil. Eleven
miRNAs (let-7g, miR-10b, miR-15a, miR-16, miR-21, miR-27a, miR-
365, miR-374b, miR-483-5p, miR-574-3p and miR-575) previously
identified in the microarray to be differentially expressed after
treatment were selected to analyze their responsiveness to dif-
ferent doses of 5-FU. Time-response data was also generated for
miR-10b, miR-21, miR-483-5p, miR-574-3p and miR-575 follow-
ing 12, 24, 36, 48, 60 and 72 h treatment. It was concluded that
miRNAs play an important regulatory role in 5-FU induced cyto-
toxicity and fit in perfectly in the intricate network of 5-FU activity
[63].
Furthermore, Koufaris et al. [64] examined the microRNA profile
in the liver of rats exposed to genotoxic (2-acetylaminofluorene)
and epigenetic (phenobarbital, diethylhexylphthalate, methapyri-
lene HCL, monuron, and chlorendic acid) chemical hepatocar-
cinogens, as well as to non-hepatocarcinogenic treatments (ben-
zophenone, and diethylthiourea). All hepatocarcinogens affects the
expression of liver mRNAs, while the hepatic microRNA profiles
are associated with the mode of action of the chemical treatments
and correspond to chemical carcinogenicity. The three nuclear
50 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57
receptor-activating chemicals (phenobarbital, benzophenone, and
diethylhexylphthalate) are characterized by the induction of the
miR-200a/200b/429. In humans, a miRNAs-Environmental associ-
ation study conducted by Xudong Wu identified 1842 concurrent
interactions between 407 miRNAs and 497 environmental chem-
icals, and thus highlighted distinct miRNAs regulated genes in
response to different environmental chemicals [65]. Since differ-
ent drugs induce various miRNAs profiles, fitting with their cellular
mode of action, the analysis of the miRNAs profile could potentially
help to categorize a potentially toxic compound.
MiRNAs are not only involved in the effects induced by an acute
exposure, but also in chronic toxicity. In the rat testis, we recently
showed that a neonatal exposure to estradiol benzoate induces
germ cells death in adulthood [66]. This phenomenon is linked
to increased levels of mir-29 which targets DNA methyl trans-
ferases and the anti-apoptotic protein MCL-1. The inhibition of
these proteins leads to the over expression of retrotransposons,
to a stimulation of apoptosis and in term to the germ cell death
[66]. However, the mechanisms of imprinting and the existence
of sensitive windows of exposure leading to irreversible miRNAs
changes remain poorly understood and represent an interesting
area for further studies.
Besides the assessment of drugs on miRNAs expression at the
individual level, a few studies showed that such alteration could
also be observed in the next generations. For instance, the offspring
of pregnant rats submitted to high fat diet presents, at the adult
age, alterations in hepatic expression of insulin like growth factor-2
and key miRNAs: miR-709, miR-122, miR-192, miR-194, miR-26a,
let-7a, let7b, let-7c, miR-494 and miR-483* [67]. Another exam-
ple is represented by the exposure to cigarette smoke. Two of the
constituents of cigarette smoke, benzo[a]pyrene and nicotine, have
been shown to induce harmful phenotypes that can be transmitted
to future generations. Indeed, in the multigenerational toxicity of
cigarette smoke, miRNAs expression alterations (mir-340, mir-365,
mir-129-3p) in the spermatozoa from human smokers and that the
altered microRNAs may be at play in pathways for maintenance
of healthy sperm and normal embryo development by control-
ling cell death/apoptosis [68]. Those studies address the potential
transgenerational effects of exposure to chemicals and additional
experiments should be realized to validate those effects. Indeed,
this field is still a matter of debate and remains under discussion.
4. miRNA as circulating biomarkers
The majority of miRNAs are intra-cellular and, as such, miRNAs
are the critical mediators in the stress cellular response, disease and
environmental stimuli [69]. Less is known, though, about extracel-
lular action of miRNAs. Stable circulating miRNAs were described
for the first time in 2008, from the plasma of patients with lym-
phoma [70]. To date, a significant number of miRNAs have been
observed outside of cells, in various body fluids. Number of miRNA
carriers have been also described such as ribonucleoprotein com-
plexes, exosomes, apoptosis bodies, membrane-derived vesicles
and high-density lipoproteins (Fig. 2) [71]. Exosomes are mem-
brane vesicles of 40–100 nm in diameter corresponding to the
internal vesicle of an endosomal compartment. It is initially formed
by the inward budding plasma membrane into multi-vesicular bod-
ies within endosomes. Exosomes and their miRNA content are
released into the extracellular compartment on the fusion of endo-
somes with the plasma membrane [72–74]. Exosomes are released
when endosomally-derived multi-vesicular bodies fuse with the
plasma membrane [75]. Apoptotic bodies and microparticles are
larger vesicles than exosomes (100–1000 nm in diameter). They
represent a heterogeneous population of vesicles that are released
by budding and blobbing of the plasma membrane and express
antigens specific of their parental cells [76,77]. Microparticles dis-
play a broad spectrum of bioactive substances and receptors on
their surface and harbor a concentrated set of cytokines, signaling
proteins, mRNAs, and microRNAs. During apoptosis, cells release
larger microparticles or apoptotic bodies that also transport spe-
cific miRNAs. Lipoproteins, particularly high-density lipoprotein
can also transport endogenous miRNAs and deliver them to recipi-
ent cells with possible functional targeting capabilities. This cellular
export is regulated by neutral sphingomyelinase and is dependent
on scavenger receptor class B type I [71]. Most of the extracel-
lular miRNAs in blood plasma and cell culture are independent
of exosomes and are bound to Argonaute 2 protein (Ago2), a
part of RNA-induced silencing complex [78]. Ago2 is the main
functional component of cytoplasmic miRNA ribonucleoprotein
complex (miRNP). Extracellular miRNAs are in the most part by-
products of dead cells that remain in extracellular space due to the
high stability of the Ago2 protein and Ago2-miRNA complex. It was
also found that extracellular circulating miRNAs could be bound to
other Ago proteins like Ago1.
These observations raised the question as to whether circulating
miRNAs may play a role, at least as potential biomarkers, in health
and disease. Indeed, circulating miRNAs are abundant in blood
and their levels in serum are stable, reproducible, and consistent
among individuals of the same species in contrast to mRNA [79,80],
which allow them to be potential non-invasive biomarkers. Several
extracellular miRNAs have now revealed specific miRNA profiles
in health and disease allowing them to represent a consistent
signature in health and some differentiating diseases [74,81,82].
For example, several tumor-specific miRNA aberrations in serum
were used as circulating biomarkers for distinguishing cancer and
cancer-free diseases (Table 2). Increasing number of reports indi-
cate that the serum miRNA expression profile can be used as a novel
serum-based biomarkers potentially offering more sensitive and
specific tests than those currently available for early diagnosis of
cancer and other diseases. Those new approaches could improve
present clinical management, including the classification of can-
cer, the prognosis estimation and the prediction of the therapeutic
efficiency [69,79,83]. A number of circulating miRNAs were used as
potential biomarkers of metabolic disorders, autoimmune, inflam-
matory diseases (Table 3).
Interestingly, the question related to miRNA as biomarkers in
health and diseases has been now extended to miRNAs as biomark-
ers in toxicology studies (Table 4). This question related to the
role of miRNA expression and epigenetic in chemical safety was
the topic of at least three recent held workshops by ILSI-HESI, US
National Academy of Sciences and ECETOC [84–88]. Indeed, that
the levels of specific circulating miRNA species may also be used to
detect and monitor the pathological development associated with
chemicals or drug-induced tissue injuries is now suggested by sev-
eral investigators [43,89–92]. Using a mouse model, the level a set
of plasma miRNAs has been associated with hepato-cellular injuries
induced by acetaminophen overdose. More specifically mir-122
and mir-192, are both enriched in the liver tissue and exhibit dose-
and exposure duration-dependent changes in the plasma that are
parallel to the serum amino-transferase levels and the histopathol-
ogy of liver degeneration [91]. MiR-103 was also reported as an
appropriate biomarker among the circulating miRNAs identified
in rats with acetaminophen-induced hepato-toxicity [43]. In the
same way, a study in human and mouse model suggested that cir-
culating miR-122 can be used as a potential novel predictive and
reliable blood marker for viral-, alcohol-, and chemical-induced
liver injury [90]. Environmental factors in daily life such as cigarette
smoking can also affect plasma miRNA profiles. Altered miRNA
expression has been reported in the smoking-related diseases [93]
and a study performed on smokers and non-smokers revealed that
plasma miRNA profiles clearly discriminate between smokers and
Table 2
Circulating miRNAs associated to cancers.

Disease miRNA Correlation Ref Disease miRNA Correlation Ref

Colorectal cancer miR-92 Differentiation of CRC from gastric [152] Gastric cancer mir-125b, Discrimination of gastric cancer from [153]
(CRC) cancer, inflammatory bowel disease, mir-199a,mir-100, healthy controls
and healthy controls let-7 g, mir-433,
mir-214
mir-21, mir-133b Discrimination of prostate cancer from [154] mir-10b, mir-21, [155]
healthy controls mir-223, mir-338,
let-7a, mir-30a-5p,
mir-126
mir-133 [156] Pancreatic cancer mir-452, mir-105, Discrimination of pancreatic cancer [157]
mir-127, mir-518-2, from healthy controls

B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57

mir-187, mir-30a-3p,
miR-196a-2
mir-221 [158] mir-18a [159]
mir-141 [160] Glioblastoma mir-21 Discrimination of glioblastoma from [161]
healthy controls

Prostate cancer miR-141 Discrimination of prostate cancer from [162] mir-128 [67]
healthy controls
mir-96 [163] Acute leukemia miR-92a decreased Differentiation between leukemia and [164]
(ratio healthy controls
miR-638/miR-92a
decreased)
mir-375, mir-141 [154] CRC and advanced miR-29a and miR-92a Discrimination of CRC and also [165]
adenome increased advanced adenoma from healthy
controls
mir-15, mir-195, [166] Diffuse large B-cell miR-155, miR-210, Differentiation of DLBCL from healthy [70]
mir-26a, let-7i lymphoma (DLBCL) miR-21 increased controls and association of miR-21
with relapse-free survival in DLBCL
Set of 15 miRNAs [167] Non-small cell lung miR- 128b, miR-152, Differentiation of NSCLC from healthy [79]
(including for cancer (NSCLC) miR-125b, miR-205, controls
instance miR-16) miR-27a, miR- 146a,
miR-222, miR-23a,
miR-24, miR-150

Breast cancer Mir-195, let7a Discrimination of breast cancer from [168] Lung cancer mir-155, let-7a Discrimination of lung cancer from [169]
healthy controls; correlation between healthy controls
miRNA levels and lymph node status
mir-145 Discrimination of breast cancer from [170] let-7a, mir-221, [171]
healthy controls mir-137, mir-372,
mir-182
mir-10b, mir-34a, [172] mir-221 and mir-222 [173]
mir-155
92a, 21 [174]

51
52 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57

Fig. 2. Mechanisms of miRNA release from cells into peripheral blood circulation and transport to target cells.
MP: microparticles; HDL: high-density lipoprotein; Ago2: Argonaute 2

Table 3
Circulating miRNAs associated to metabolic disorders, auto-immune and inflammatory diseases.

Physiopathological status miRNAs Correlation References

Pregnancy mir-527, miR-520d-5p and miR-526a Discrimination of pregnant women from non-pregnant women [175]
Non-small-cell lung carcinoma let-7f, miR-20b and miR-30e-3p Discrimination between NSCLC and healthy control [176]
(NSCLC)
Rheumatoid arthritis miR-132 decreased Distinction between RA or OA [177]
(RA)/osteoarthritis (OA)
mir-16 and miR-146a Correlation with tender joint counts and 28-joint Disease
Activity Score

Coronary artery disease (CAD) miR-126, miR-17, miR-92a, miR-155, miR-145 Distinction between CAD and healthy controls [178]
miR-133a and miR-208a

Chron disease miR-149 [179]

Diabetes miR-144 Increased [180]
Alzheimer’s disease miR-137, 181c, 9, 29a/b Reduced [181]
Duchenne muscular dystrophy miR-1, 133a, 206 Increased [182]

non-smokers. This observation suggested that repeated cigarette
smoking substantially alters the plasma miRNA profile [94]. Expo-
sure to metal-rich particulate matter has been reported to modify
the expression of candidate miRNAs in peripheral blood leukocytes
of workers at an electric-furnace steel plant. Indeed, expression of
miRNA-222 and miRNA-21 was significantly upregulated in post-
exposure samples [95].
Circulating miRNAs may also represent good markers for
hypertension-induced heart failure and for the response to thera-
peutic treatment. Indeed, in a rat model, miRNAs arrays on plasma

Table 4
MiRNA expression in seminal plasma from infertile men.

Diseases miRNAs Correlation References

Oligoastheno-zoospermia
miR-141, miR-29a, miR-429, Upregulated in spermatozoa samples from oligoasthenozoospermic [183]
miR200a patients compared with those from normozoospermic
miR- 34b, miR-19a, miR-16, Downregulated in supernatant sperm samples from
miR-122 oligoasthenozoospermic patients compared with those from
normozoospermic

Astheno-zoospermia miR-30a, miR-26a, miR-200a,
miR-141, miR-29a, miR-24
miR-1973, miR-34b, miR-122
Upregulated in supernatant sperm samples from asthenozoospermic
patients compared with those from normozoospermic
Downregulated in supernatant sperm samples from
asthenozoospermic patients compared with those from
normozoospermic

Infertility miR-574-5p, miR-297, Upregulated in the semen of infertile males with semen abnormalities [184]
miR-122, miR-1275, miR-373,
miR-185 and miR-193b
miR-100, miR-512-3p, miR-16, Downregulated in the semen of infertile males with semen
miR-19b, miR-23b and abnormalities
miR-26a

Non obstructive miR-19b and let-7a Upregulated in supernatant sperm from idiopathic infertile males with [185]
azoospermia non obstructive azoospermia compared with fertile controls
 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57
highlighted modifications in miR-16, miR-20b, miR-93, miR-106b,
miR-223, and miR-423-5p. A time course study showed that those
changes are correlated to disease progression [96].
In kidney diseases, a number of studies have highlighted the sen-
sitivity of miRNAs detection in plasma and urine (for review [97]).
In injury induced by renal ischemia reperfusion or streptozotocin
induced diabetes in mice, miR-10a and miR-30d concentrations
in plasma and urine are positively correlated with the degree of
kidney injury [98]. In humans, urinary and plasma miR-21 are asso-
ciated with severe acute kidney injury [99].
Together, there are now many studies establishing a potential
link between the level of circulating miRNA and clinical diagnosis
or prognosis in many pathologies or injuries. The use of circulating
miRNAs in body fluids as potential toxicological biomarkers, and
the link between miRNA-related pharmaco-genomics and adverse
drug reactions will be a matter of future investigations [43].
5. Conclusions
MiRNAs are highly conserved between human and animal mod-
els and can regulate biological pathways by repressing target
proteins. Chemicals can affect those pathways by inducing their
down or up-regulation. Also, miRNA profiling in response to toxic
compounds can provide toxicant-specific profiles in specific organs.
Given their specificity to tested compounds, the time and dose
dependency of their modifications, the sensitivity and simplicity
of their detection in various tissues, miRNAs appear like promis-
ing tools in toxicological studies. Interestingly, circulating miRNAs
could be accessible through non-invasive protocols and stable. Cir-
culating miRNAs thus represent ideal candidates in toxicological
studies, and may be used as biomarkers of chemical exposure for
safety assessment.
However, with regard to the involvement of miRNA expres-
sion in the effects of exposure to chemicals, there are at least two
main observations that should be taken into account. Firstly, while
there are a growing number of reports showing altered miRNAs
profile following exposure to drugs/chemicals, the mechanisms
leading to the pathological phenotype remain poorly understood.
One of the major limitations of numerous studies analyzing the
miRNAs expression changes in experimental models after exposure
to chemicals is that these changes, generally, are viewed associ-
ated rather than causal to the adverse apical endpoints. One of the
appropriate approaches to establish a causal link between miRNA
expression and the adverse effects could be the following; iden-
tify first the adverse effect and then try to delineate and dissect
the supporting miRNA expression changes. Recently, by using such
an approach, alterations in mir-29 family members that induced
adult germ cell death process leading to infertility were identified
following neonatal exposure to the xenoestrogen estradiol ben-
zoate [66]. Secondly, a number of studies have highlighted the dose
and time dependency of miRNAs profile after exposure to chemi-
cal compounds. Furthermore, studies have showed that exposure
can lead to long term changes, and possibly irreversible. The irre-
versibility of miRNA expression changes is likely occurring when
the exposure takes place during critical developmental periods i.e.
the in utero and perinatal periods that are specifically sensitive to
chemicals. Those observations are to be viewed in the context of
the concept of Developmental Origin of Health and Diseases (also
termed the Barker’s hypothesis). In this context, low-dose effects
have received considerable attention from the scientific and regula-
tory communities. Analysis of miRNAs seems to be an appropriate
approach in risk assessment at low doses, and long term effects
induced by EDCs. Based on the increasing amounts of accumu-
lating data on miRNA expression alterations following exposure
to chemicals and the fact that circulating miRNAs appear as very
53
interesting biomarkers, the possibility that miRNAs measurement
could be included in toxicological studies is open. While the field of
epigenetic (including miRNA but also methylation and chromatin
remodeling) is a rapidly growing field in both understanding of
these mechanisms in health and diseases as well as in terms of
availability of tools, some key questions remain still to be clarified:
(i) are miRNA expression changes causes or consequences of the
adverse effects? (ii) Are they stable and irreversible? (86).
Conflict of interest
None.
References
[1] V.N. Kim, J. Han, M.C. Siomi, Biogenesis of small RNAs in animals, Nat. Rev.
Mol. Cell Biol. 10 (2009) 126–139.
[2] M. Ghildiyal, P.D. Zamore, Small silencing RNAs: an expanding universe, Nat.
Rev. Genet. 10 (2009) 94–108.
[3] C.D. Malone, G.J. Hannon, Small RNAs as guardians of the genome, Cell 136
(2009) 656–668.
[4] J.C. Mathers, G. Strathdee, C.L. Relton, Induction of epigenetic alterations by
dietary and other environmental factors, Adv. Genet. 71 (2010) 3–39.
[5] B.P. Lewis, C.B. Burge, D.P. Bartel, Conserved seed pairing, often flanked by
adenosines, indicates that thousands of human genes are microRNA targets,
Cell 120 (2005) 15–20.
[6] M. Szyf, The dynamic epigenome and its implications in toxicology, Toxicol.
Sci. 100 (2007) 7–23.
[7] Y. Saito, P.A. Jones, Epigenetic activation of tumor suppressor microRNAs in
human cancer cells, Cell Cycle 5 (2006) 2220–2222.
[8] L. Meunier, B. Siddeek, A. Vega, N. Lakhdari, L. Inoubli, R.P. Bellon, G. Lemaire,
C. Mauduit, M. Benahmed, Perinatal programming of adult rat germ cell death
after exposure to xenoestrogens: role of microRNA miR-29 family in the
down-regulation of DNA methyltransferases and Mcl-1, Endocrinology 153
(2012) 1936–1947.
[9] M.J. LeBaron, R.J. Rasoulpour, J. Klapacz, R.G. Ellis-Hutchings, H.M. Hollnagel,
B.B. Gollapudi, Epigenetics and chemical safety assessment, Mutat. Res. 705
(2010) 83–95.
[10] M.R. Fabian, N. Sonenberg, The mechanics of miRNA-mediated gene silencing:
a look under the hood of miRISC, Nat. Struct. Mol. Biol. 19 (2012) 586–593.
[11] S.D. Fiedler, M.Z. Carletti, L.K. Christenson, Quantitative RT-PCR methods for
mature microRNA expression analysis, Methods Mol. Biol. 630 (2010) 49–64.
[12] E. Varallyay, J. Burgyan, Z. Havelda, MicroRNA detection by northern blotting
using locked nucleic acid probes, Nat. Protoc. 3 (2008) 190–196.
[13] S. Ambs, R.L. Prueitt, M. Yi, R.S. Hudson, T.M. Howe, F. Petrocca, T.A. Wallace,
C.G. Liu, S. Volinia, G.A. Calin, H.G. Yfantis, R.M. Stephens, C.M. Croce, Genomic
profiling of microRNA and messenger RNA reveals deregulated microRNA
expression in prostate cancer, Cancer Res. 68 (2008) 6162–6170.
[14] Z. Williams, I.Z. Ben-Dov, R. Elias, A. Mihailovic, M. Brown, Z. Rosenwaks,
T. Tuschl, Comprehensive profiling of circulating microRNA via small RNA
sequencing of cDNA libraries reveals biomarker potential and limitations,
Proc. Natl. Acad. Sci. U.S.A. 110 (2013) 4255–4260.
[15] A.M. Monteys, R.M. Spengler, J. Wan, L. Tecedor, K.A. Lennox, Y. Xing, B.L.
Davidson, Structure and activity of putative intronic miRNA promoters, RNA
16 (2013) 495–505.
[16] Y. Lee, M. Kim, J. Han, K.H. Yeom, S. Lee, S.H. Baek, V.N. Kim, MicroRNA genes
are transcribed by RNA polymerase II, EMBO J. 23 (2004) 4051–4060.
[17] G.M. Borchert, W. Lanier, B.L. Davidson, RNA polymerase III transcribes human
microRNAs, Nat. Struct. Mol. Biol. 13 (2006) 1097–1101.
[18] Y.S. Lee, A. Dutta, MicroRNAs in cancer, Annu. Rev. Pathol. 4 (2009) 199–227.
[19] F. Ozsolak, L.L. Poling, Z. Wang, H. Liu, X.S. Liu, R.G. Roeder, X. Zhang, J.S. Song,
D.E. Fisher, Chromatin structure analyses identify miRNA promoters, Genes
Dev. 22 (2008) 3172–3183.
[20] V. Libri, P. Miesen, R.P. van Rij, A.H. Buck, Regulation of microRNA biogene-
sis and turnover by animals and their viruses, Cell. Mol. Life Sci. 70 (2013)
3525–3544.
[21] Y. Lee, C. Ahn, J. Han, H. Choi, J. Kim, J. Yim, J. Lee, P. Provost, O. Radmark, S.
Kim, V.N. Kim, The nuclear RNase III Drosha initiates microRNA processing,
Nature 425 (2003) 415–419.
[22] R.I. Gregory, K.P. Yan, G. Amuthan, T. Chendrimada, B. Doratotaj, N. Cooch, R.
Shiekhattar, The Microprocessor complex mediates the genesis of microRNAs,
Nature 432 (2004) 235–240.
[23] R. Triboulet, H.M. Chang, R.J. Lapierre, R.I. Gregory, Post-transcriptional con-
trol of DGCR8 expression by the microprocessor, RNA 15 (2009) 1005–1011.
[24] S. Kadener, J. Rodriguez, K.C. Abruzzi, Y.L. Khodor, K. Sugino, M.T. Marr 2nd,
S. Nelson, M. Rosbash, Genome-wide identification of targets of the drosha-
pasha/DGCR8 complex, RNA 15 (2009) 537–545.
[25] E. Bernstein, A.A. Caudy, S.M. Hammond, G.J. Hannon, Role for a bidentate
ribonuclease in the initiation step of RNA interference, Nature 409 (2001)
363–366.
54 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57
[26] G. Hutvagner, J. McLachlan, A.E. Pasquinelli, E. Balint, T. Tuschl, P.D. Zamore,
A cellular function for the RNA-interference enzyme Dicer in the maturation
of the let-7 small temporal RNA, Science 293 (2001) 834–838.
[27] D.P. Bartel, MicroRNAs: target recognition and regulatory functions, Cell 136
(2009) 215–233.
[28] J. Wang, M. Lu, C. Qiu, Q. Cui, TransmiR: a transcription factor-microRNA
regulation database, Nucleic Acids Res. 38 (2010) D119–D122.
[29] B.N. Davis-Dusenbery, A. Hata, Smad-mediated miRNA processing: a critical
role for a conserved RNA sequence, RNA Biol. 8 (2011) 71–76.
[30] X. Wang, X. Zhao, P. Gao, M. Wu, c-Myc modulates microRNA processing via
the transcriptional regulation of Drosha, Sci. Rep. 3 (2013) 1942.
[31] H.I. Suzuki, K. Yamagata, K. Sugimoto, T. Iwamoto, S. Kato, K. Miyazono, Mod-
ulation of microRNA processing by p53, Nature 460 (2009) 529–533.
[32] S. Hong, H. Noh, H. Chen, R. Padia, Z.K. Pan, S.B. Su, Q. Jing, H.F. Ding, S. Huang,
Signaling by p38 MAPK stimulates nuclear localization of the microprocessor
component p68 for processing of selected primary microRNAs, Sci. Signal. 6
(2013) ra16.
[33] S. Kawai, A. Amano, BRCA1 regulates microRNA biogenesis via the DROSHA
microprocessor complex, J. Cell Biol. 197 (2012) 201–208.
[34] K. Yamagata, S. Fujiyama, S. Ito, T. Ueda, T. Murata, M. Naitou, K. Takeyama,
Y. Minami, B.W. O’Malley, S. Kato, Maturation of microRNA is hormonally
regulated by a nuclear receptor, Mol. Cell 36 (2009) 340–347.
[35] S. Sakamoto, K. Aoki, T. Higuchi, H. Todaka, K. Morisawa, N. Tamaki, E. Hatano,
A. Fukushima, T. Taniguchi, Y. Agata, The NF90-NF45 complex functions as a
negative regulator in the microRNA processing pathway, Mol. Cell. Biol. 29
(2009) 3754–3769.
[36] M.A. Newman, J.M. Thomson, S.M. Hammond, Lin-28 interaction with the Let-
7 precursor loop mediates regulated microRNA processing, RNA 14 (2008)
1539–1549.
[37] E. Piskounova, C. Polytarchou, J.E. Thornton, R.J. LaPierre, C. Pothoulakis, J.P.
Hagan, D. Iliopoulos, R.I. Gregory, Lin28A and Lin28B inhibit let-7 microRNA
biogenesis by distinct mechanisms, Cell 147 (2011) 1066–1079.
[38] G. Michlewski, J.F. Caceres, Antagonistic role of hnRNP A1 and KSRP in the
regulation of let-7a biogenesis, Nat. Struct. Mol. Biol. 17 (2010) 1011–1018.
[39] N.R. Choudhury, G. Michlewski, Terminal loop-mediated control of microRNA
biogenesis, Biochem. Soc. Trans. 40 (2012) 789–793.
[40] S. Bail, M. Swerdel, H. Liu, X. Jiao, L.A. Goff, R.P. Hart, M. Kiledjian, Differential
regulation of microRNA stability, RNA 16 (2006) 1032–1039.
[41] M.P. Gantier, C.E. McCoy, I. Rusinova, D. Saulep, D. Wang, D. Xu, A.T. Irving, M.A.
Behlke, P.J. Hertzog, F. Mackay, B.R. Williams, Analysis of microRNA turnover
in mammalian cells following Dicer1 ablation, Nucleic Acids Res. 39 (2011)
5692–5703.
[42] J.O. Westholm, E. Ladewig, K. Okamura, N. Robine, E.C. Lai, Common and dis-
tinct patterns of terminal modifications to mirtrons and canonical microRNAs,
RNA 18 (2012) 177–192.
[43] T. Yokoi, M. Nakajima, microRNAs as mediators of drug toxicity, Annu. Rev.
Pharmacol. Toxicol. 53 (2013) 377–400.
[44] J.K. Rieger, K. Klein, S. Winter, U.M. Zanger, Expression variability of absorp-
tion, distribution, metabolism, excretion-related microRNAs in human liver:
influence of nongenetic factors and association with gene expression, Drug
Metab. Dispos. 41 (2013) 1752–1762.
[45] S. Takagi, M. Nakajima, T. Mohri, T. Yokoi, Post-transcriptional regulation
of human pregnane X receptor by micro-RNA affects the expression of
cytochrome P450 3A4, J. Biol. Chem. 283 (2008) 9674–9680.
[46] W. Hu, J. Zhao, G. Pei, Activation of aryl hydrocarbon receptor (AhR) by
Tranilast, an anti-allergy drug, promotes miR-302 expression and cell repro-
gramming, J. Biol. Chem. 288 (2013) 22972–22984.
[47] R. Shizu, S. Benoki, Y. Numakura, S. Kodama, M. Miyata, Y. Yamazoe,
K. Yoshinari, Xenobiotic-induced hepatocyte proliferation associated with
constitutive active/androstane receptor (CAR) or peroxisome proliferator-
activated receptor alpha (PPARalpha) is enhanced by pregnane X receptor
(PXR) activation in mice, PLoS One 8 (2013) e61802.
[48] K. Kida, M. Nakajima, T. Mohri, Y. Oda, S. Takagi, T. Fukami, T. Yokoi, PPARalpha
is regulated by miR-21 and miR-27b in human liver, Pharm. Res. 28 (2011)
2467–2476.
[49] L. Zheng, G.C. Lv, J. Sheng, Y.D. Yang, Effect of miRNA-10b in regulating cellular
steatosis level by targeting PPAR-alpha expression, a novel mechanism for the
pathogenesis of NAFLD, J. Gastroenterol. Hepatol. 25 (2010) 156–163.
[50] Y.M. Shah, K. Morimura, Q. Yang, T. Tanabe, M. Takagi, F.J. Gonzalez, Perox-
isome proliferator-activated receptor alpha regulates a microRNA-mediated
signaling cascade responsible for hepatocellular proliferation, Mol. Cell. Biol.
27 (2007) 4238–4247.
[51] T. Nakahama, H. Hanieh, N.T. Nguyen, I. Chinen, B. Ripley, D. Millrine, S.
Lee, K.K. Nyati, P.K. Dubey, K. Chowdhury, Y. Kawahara, T. Kishimoto, Aryl
hydrocarbon receptor-mediated induction of the microRNA-132/212 cluster
promotes interleukin-17-producing T-helper cell differentiation, Proc. Natl.
Acad. Sci. U.S.A. 110 (2013) 11964–11969.
[52] B.D. Adams, H. Furneaux, B.A. White, The micro-ribonucleic acid (miRNA)
miR-206 targets the human estrogen receptor-alpha (ERalpha) and represses
ERalpha messenger RNA and protein expression in breast cancer cell lines,
Mol. Endocrinol. 21 (2007) 1132–1147.
[53] J.J. Zhao, J. Lin, H. Yang, W. Kong, L. He, X. Ma, D. Coppola, J.Q. Cheng, MicroRNA-
221/222 negatively regulates estrogen receptor alpha and is associated with
tamoxifen resistance in breast cancer, J. Biol. Chem. 283 (2008) 31079–31086.
[54] J. Xiong, D. Yu, N. Wei, H. Fu, T. Cai, Y. Huang, C. Wu, X. Zheng, Q. Du,
D. Lin, Z. Liang, An estrogen receptor alpha suppressor, microRNA-22, is
downregulated in estrogen receptor alpha-positive human breast cancer cell
lines and clinical samples, FEBS J. 277 (2010) 1684–1694.
[55] Z. Yang, L. Wang, Regulation of microRNA expression and function by nuclear
receptor signaling, Cell Biosci. 1 (2011) 31.
[56] J. Wu, J. Bao, L. Wang, Y. Hu, C. Xu, MicroRNA-184 downregulates nuclear
receptor corepressor 2 in mouse spermatogenesis, BMC Dev. Biol. 11 (2011)
64.
[57] P. Landgraf, M. Rusu, R. Sheridan, A. Sewer, N. Iovino, A. Aravin, S. Pfef-
fer, A. Rice, A.O. Kamphorst, M. Landthaler, C. Lin, N.D. Socci, L. Hermida,
V. Fulci, S. Chiaretti, R. Foa, J. Schliwka, U. Fuchs, A. Novosel, R.U. Muller,
B. Schermer, U. Bissels, J. Inman, Q. Phan, M. Chien, D.B. Weir, R. Choksi,
G. De Vita, D. Frezzetti, H.I. Trompeter, V. Hornung, G. Teng, G. Hartmann,
M. Palkovits, R. Di Lauro, P. Wernet, G. Macino, C.E. Rogler, J.W. Nagle, J. Ju,
F.N. Papavasiliou, T. Benzing, P. Lichter, W. Tam, M.J. Brownstein, A. Bosio, A.
Borkhardt, J.J. Russo, C. Sander, M. Zavolan, T. Tuschl, A mammalian microRNA
expression atlas based on small RNA library sequencing, Cell 129 (2007)
1401–1414.
[58] Y. Wang, T. Weng, D. Gou, Z. Chen, N.R. Chintagari, L. Liu, Identification of rat
lung-specific microRNAs by micoRNA microarray: valuable discoveries for
the facilitation of lung research, BMC Genomics 8 (2007) 29.
[59] M. Satoh, Y. Minami, Y. Takahashi, T. Tabuchi, M. Nakamura, Expression of
microRNA-208 is associated with adverse clinical outcomes in human dilated
cardiomyopathy, J. Card. Fail. 16 (2010) 404–410.
[60] C.E. Grueter, E. van Rooij, B.A. Johnson, S.M. DeLeon, L.B. Sutherland, X. Qi, L.
Gautron, J.K. Elmquist, R. Bassel-Duby, E.N. Olson, A cardiac microRNA gov-
erns systemic energy homeostasis by regulation of MED13, Cell 149 (2012)
671–683.
[61] M.D. Shapiro, J. Bagley, J. Latz, J.G. Godwin, X. Ge, S.G. Tullius, J. Iacomini,
MicroRNA expression data reveals a signature of kidney damage following
ischemia reperfusion injury, PLoS One 6 (2011) e23011.
[62] C. Vacchi-Suzzi, Y. Bauer, B.R. Berridge, S. Bongiovanni, K. Gerrish, H.K.
Hamadeh, M. Letzkus, J. Lyon, J. Moggs, R.S. Paules, F. Pognan, F. Staedtler,
M.P. Vidgeon-Hart, O. Grenet, P. Couttet, Perturbation of microRNAs in rat
heart during chronic doxorubicin treatment, PLoS One 7 (2012) e40395.
[63] M.Y. Shah, X. Pan, L.N. Fix, M.A. Farwell, B. Zhang, 5-Fluorouracil drug alters
the microRNA expression profiles in MCF-7 breast cancer cells, J. Cell. Physiol.
226 (2011) 1868–1878.
[64] C. Koufaris, J. Wright, R.A. Currie, N.J. Gooderham, Hepatic microRNA pro-
files offer predictive and mechanistic insights after exposure to genotoxic
and epigenetic hepatocarcinogens, Toxicol. Sci. 128 (2012) 532–543.
[65] X. Wu, Y. Song, Preferential regulation of miRNA targets by environmental
chemicals in the human genome, BMC Genomics 12 (2011) 244.
[66] L. Meunier, B. Siddeek, A. Vega, N. Lakhdari, L. Inoubli, R.P. Bellon, G. Lemaire, C.
Mauduit, M. Benahmed, Perinatal programming of adult rat germ cell death
after exposure to xenoestrogens: role of microRNA miR -29 family in the
down-regulation of DNA methyltransferases and Mcl-1, Endocrinology 153
(2012) 1936–1947.
[67] Y. Zhang, T. Chao, R. Li, W. Liu, Y. Chen, X. Yan, Y. Gong, B. Yin, B. Qiang, J.
Zhao, J. Yuan, X. Peng, MicroRNA-128 inhibits glioma cells proliferation by
targeting transcription factor E2F3a, J. Mol. Med. (Berl.) 87 (2009) 43–51.
[68] E.L. Marczylo, A.A. Amoako, J.C. Konje, T.W. Gant, T.H. Marczylo, Smoking
induces differential miRNA expression in human spermatozoa: a potential
transgenerational epigenetic concern? Epigenetics 7 (2012) 432–439.
[69] J.T. Mendell, E.N. Olson, MicroRNAs in stress signaling and human disease,
Cell 148 (2012) 1172–1187.
[70] C.H. Lawrie, S. Gal, H.M. Dunlop, B. Pushkaran, A.P. Liggins, K. Pulford, A.H.
Banham, F. Pezzella, J. Boultwood, J.S. Wainscoat, C.S. Hatton, A.L. Harris,
Detection of elevated levels of tumour-associated microRNAs in serum of
patients with diffuse large B-cell lymphoma, Br. J. Haematol. 141 (2008)
672–675.
[71] K.C. Vickers, B.T. Palmisano, B.M. Shoucri, R.D. Shamburek, A.T. Remaley,
MicroRNAs are transported in plasma and delivered to recipient cells by
high-density lipoproteins, Nat. Cell. Biol. 13 (2011) 423–433.
[72] C. Thery, L. Zitvogel, S. Amigorena, Exosomes: composition, biogenesis and
function, Nat. Rev. Immunol. 2 (2002) 569–579.
[73] W. Stoorvogel, M.J. Kleijmeer, H.J. Geuze, G. Raposo, The biogenesis and func-
tions of exosomes, Traffic 3 (2002) 321–330.
[74] R.A. Boon, K.C. Vickers, Intercellular transport of microRNAs, Arterioscler.
Thromb. Vasc. Biol. 33 (2013) 186–192.
[75] A. Allegra, A. Alonci, S. Campo, G. Penna, A. Petrungaro, D. Gerace, C. Musolino,
Circulating microRNAs: new biomarkers in diagnosis, prognosis and treat-
ment of cancer (review), Int. J. Oncol. 41 (2012) 1897–1912.
[76] L. Xu, B.F. Yang, J. Ai, MicroRNA transport: a new way in cell communication,
J. Cell. Physiol. 228 (2013) 1713–1719.
[77] S.F. Mause, C. Weber, Microparticles: protagonists of a novel communica-
tion network for intercellular information exchange, Circ. Res. 107 (2010)
1047–1057.
[78] A. Turchinovich, L. Weiz, A. Langheinz, B. Burwinkel, Characterization of extra-
cellular circulating microRNA, Nucleic Acids Res. 39 (2011) 7223–7233.
[79] X. Chen, Y. Ba, L. Ma, X. Cai, Y. Yin, K. Wang, J. Guo, Y. Zhang, J. Chen, X. Guo,
Q. Li, X. Li, W. Wang, J. Wang, X. Jiang, Y. Xiang, C. Xu, P. Zheng, J. Zhang, R. Li,
H. Zhang, X. Shang, T. Gong, G. Ning, K. Zen, C.Y. Zhang, Characterization of
microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and
other diseases, Cell Res. 18 (2008) 997–1006.
[80] M.P. Hunter, N. Ismail, X. Zhang, B.D. Aguda, E.J. Lee, L. Yu, T. Xiao, J. Schafer,
M.L. Lee, T.D. Schmittgen, S.P. Nana-Sinkam, D. Jarjoura, C.B. Marsh, Detection
 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57
of microRNA expression in human peripheral blood microvesicles, PLoS One
3 (2008) e3694.
[81] N. Scholer, C. Langer, F. Kuchenbauer, Circulating microRNAs as biomarkers
– true Blood? Genome Med. 3 (2011) 72.
[82] A. Keller, P. Leidinger, A. Bauer, A. Elsharawy, J. Haas, C. Backes, A. Wendschlag,
N. Giese, C. Tjaden, K. Ott, J. Werner, T. Hackert, K. Ruprecht, H. Huwer, J. Hue-
bers, G. Jacobs, P. Rosenstiel, H. Dommisch, A. Schaefer, J. Muller-Quernheim,
B. Wullich, B. Keck, N. Graf, J. Reichrath, B. Vogel, A. Nebel, S.U. Jager, P.
Staehler, I. Amarantos, V. Boisguerin, C. Staehler, M. Beier, M. Scheffler, M.W.
Buchler, J. Wischhusen, S.F. Haeusler, J. Dietl, S. Hofmann, H.P. Lenhof, S.
Schreiber, H.A. Katus, W. Rottbauer, B. Meder, J.D. Hoheisel, A. Franke, E.
Meese, Toward the blood-borne miRNome of human diseases, Nat. Methods
8 (2011) 841–843.
[83] J.A. Weber, D.H. Baxter, S. Zhang, D.Y. Huang, K.H. Huang, M.J. Lee, D.J. Galas,
K. Wang, The microRNA spectrum in 12 body fluids, Clin. Chem. 56 (2010)
1733–1741.
[84] E. Buckley, Epigenetic effects of chemicals not ready for regulatory “prime-
time”, Pestic. Toxic Chem. News 37 (2009), http://www.agra-net.com/aius/
home.jsp?pagetitle=aiusfp&pubId=ag100
[85] J.I. Goodman, K.A. Augustine, M.L. Cunnningham, D. Dixon, Y.P. Dragan, J.G.
Falls, R.J. Rasoulpour, R.C. Sills, R.D. Storer, D.C. Wolf, S.D. Pettit, What do we
need to know prior to thinking about incorporating an epigenetic evaluation
into safety assessments? Toxicol. Sci. 116 (2010) 375–381.
[86] B. Hileman, Chemicals can turn genes on and off, Environ. Health News 3
(August) (2009).
[87] ECETOC WR 23 - Epigenetics and Chemical Safety Rome, 2011.
[88] C.C. Priestley, M. Anderton, A.T. Doherty, P. Duffy, H.R. Mellor, H. Powell, R.
Roberts, Epigenetics – relevance to drug safety science, Toxicol. Res. 1 (2012)
23–31.
[89] L. Hou, X. Zhang, D. Wang, A. Baccarelli, Environmental chemical exposures
and human epigenetics, Int. J. Epidemiol. 41 (2012) 79–105.
[90] Y. Zhang, Y. Jia, R. Zheng, Y. Guo, Y. Wang, H. Guo, M. Fei, S. Sun, Plasma
microRNA-122 as a biomarker for viral-, alcohol-, and chemical-related hep-
atic diseases, Clin. Chem. 56 (2010) 1830–1838.
[91] K. Wang, S. Zhang, B. Marzolf, P. Troisch, A. Brightman, Z. Hu, L.E. Hood, D.J.
Galas, Circulating microRNAs, potential biomarkers for drug-induced liver
injury, Proc. Natl. Acad. Sci. U.S.A. 106 (2009) 4402–4407.
[92] T. Yokoi, M. Nakajima, Toxicological implications of modulation of gene
expression by microRNAs, Toxicol. Sci. 123 (2011) 1–14.
[93] A. Banerjee, K. Luettich, MicroRNAs as potential biomarkers of smoking-
related diseases, Biomark Med. 6 (2012) 671–684.
[94] K. Takahashi, S. Yokota, N. Tatsumi, T. Fukami, T. Yokoi, M. Nakajima, Cigarette
smoking substantially alters plasma microRNA profiles in healthy subjects,
Toxicol. Appl. Pharmacol. 272 (2013) 154–160.
[95] V. Bollati, B. Marinelli, P. Apostoli, M. Bonzini, F. Nordio, M. Hoxha, V. Pego-
raro, V. Motta, L. Tarantini, L. Cantone, J. Schwartz, P.A. Bertazzi, A. Baccarelli,
Exposure to metal-rich particulate matter modifies the expression of candi-
date microRNAs in peripheral blood leukocytes, Environ. Health Perspect. 118
(2010) 763–768.
[96] B.A. Dickinson, H.M. Semus, R.L. Montgomery, C. Stack, P.A. Latimer,
S.M. Lewton, J.M. Lynch, T.G. Hullinger, A.G. Seto, E. van Rooij, Plasma
microRNAs serve as biomarkers of therapeutic efficacy and disease pro-
gression in hypertension-induced heart failure, Eur. J. Heart Fail. 15 (2010)
650–659.
[97] J.M. Lorenzen, T. Thum, Circulating and urinary microRNAs in kidney disease,
Clin. J. Am. Soc. Nephrol. 7 (2012) 1528–1533.
[98] N. Wang, Y. Zhou, L. Jiang, D. Li, J. Yang, C.Y. Zhang, K. Zen, Urinary microRNA-
10a and microRNA-30d serve as novel, sensitive and specific biomarkers for
kidney injury, PloS One 7 (2012) e51140.
[99] J. Du, X. Cao, L. Zou, Y. Chen, J. Guo, Z. Chen, S. Hu, Z. Zheng, MicroRNA-21
and risk of severe acute kidney injury and poor outcomes after adult cardiac
surgery, PloS One 8 (2013) e63390.
[100] F. Wang, C. Li, W. Liu, Y. Jin, Effect of exposure to volatile organic compounds
(VOCs) on airway inflammatory response in mice, J. Toxicol. Sci. 37 (2012)
739–748.
[101] J. Bolleyn, J. Fraczek, M. Vinken, D. Lizarraga, S. Gaj, J.H. van Delft,
V. Rogiers, T. Vanhaecke, Effect of Trichostatin A on miRNA expres-
sion in cultures of primary rat hepatocytes, Toxicol. In Vitro 25 (2011)
1173–1182.
[102] C.J. Marsit, K. Eddy, K.T. Kelsey, MicroRNA responses to cellular stress, Cancer
Res. 66 (2006) 10843–10848.
[103] Y. Cao, S.L. Yu, Y. Wang, G.Y. Guo, Q. Ding, R.H. An, MicroRNA-dependent
regulation of PTEN after arsenic trioxide treatment in bladder cancer cell line
T24, Tumour Biol. 32 (2011) 179–188.
[104] A.I. Pogue, Y.Y. Li, J.G. Cui, Y. Zhao, T.P. Kruck, M.E. Percy, M.A. Tarr, W.J. Lukiw,
Characterization of an NF-kappaB-regulated, miRNA-146a-mediated down-
regulation of complement factor H (CFH) in metal-sulfate-stressed human
brain cells, J. Inorg. Biochem. 103 (2009) 1591–1595.
[105] W.J. Lukiw, A.I. Pogue, Induction of specific micro RNA (miRNA) species by
ROS-generating metal sulfates in primary human brain cells, J. Inorg. Biochem.
101 (2007) 1265–1269.
[106] V. Bollati, B. Marinelli, P. Apostoli, M. Bonzini, F. Nordio, M. Hoxha, V. Pego-
raro, V. Motta, L. Tarantini, L. Cantone, J. Schwartz, P.A. Bertazzi, A. Baccarelli,
Exposure to metal-rich particulate matter modifies the expression of can-
didate microRNAs in peripheral blood leukocytes, Environ. Health. Perspect.
118 (2010) 763–768.
55
[107] A. Izzotti, G.A. Calin, P. Arrigo, V.E. Steele, C.M. Croce, S. De Flora, Downregula-
tion of microRNA expression in the lungs of rats exposed to cigarette smoke,
FASEB J. 23 (2009) 806–812.
[108] F. Schembri, S. Sridhar, C. Perdomo, A.M. Gustafson, X. Zhang, A. Ergun, J. Lu, G.
Liu, X. Zhang, J. Bowers, C. Vaziri, K. Ott, K. Sensinger, J.J. Collins, J.S. Brody, R.
Getts, M.E. Lenburg, A. Spira, MicroRNAs as modulators of smoking-induced
gene expression changes in human airway epithelium, Proc. Natl. Acad. Sci.
U.S.A. 106 (2009) 2319–2324.
[109] S. Kalscheuer, X. Zhang, Y. Zeng, P. Upadhyaya, Differential expression of
microRNAs in early-stage neoplastic transformation in the lungs of F344 rats
chronically treated with the tobacco carcinogen 4-(methylnitrosamino)-1-
(3-pyridyl)-1-butanone, Carcinogenesis 29 (2008) 2394–2399.
[110] B. Zhang, X. Pan, RDX induces aberrant expression of microRNAs in mouse
brain and liver, Environ. Health Perspect. 117 (2009) 231–240.
[111] T. Fukushima, Y. Hamada, H. Yamada, I. Horii, Changes of micro-RNA expres-
sion in rat liver treated by acetaminophen or carbon tetrachloride–regulating
role of micro-RNA for RNA expression, J. Toxicol. Sci. 32 (2007) 401–409.
[112] E. Elyakim, E. Sitbon, A. Faerman, S. Tabak, E. Montia, L. Belanis, A. Dov, E.G.
Marcusson, C.F. Bennett, A. Chajut, D. Cohen, N. Yerushalmi, hsa-miR-191 is a
candidate oncogene target for hepatocellular carcinoma therapy, Cancer Res.
70 (2010) 8077–8087.
[113] M. Avissar-Whiting, K.R. Veiga, K.M. Uhl, M.A. Maccani, L.A. Gagne, E.L. Moen,
C.J. Marsit, A. Bisphenol, exposure leads to specific microRNA alterations in
placental cells, Reprod. Toxicol. 29 (2010) 401–406.
[114] J.S. Lee, D. Semela, J. Iredale, V.H. Shah, Sinusoidal remodeling and angiogen-
esis: a new function for the liver-specific pericyte? Hepatology (Baltimore,
Md.) 45 (2007) 817–825.
[115] P. Sathyan, H.B. Golden, R.C. Miranda, Competing interactions between micro-
RNAs determine neural progenitor survival and proliferation after ethanol
exposure: evidence from an ex vivo model of the fetal cerebral cortical neu-
roepithelium, J. Neurosci. 27 (2007) 8546–8557.
[116] I.P. Pogribny, V.P. Tryndyak, A. Boyko, R. Rodriguez-Juarez, F.A. Beland, O.
Kovalchuk, Induction of microRNAome deregulation in rat liver by long-term
tamoxifen exposure, Mutat. Res. 619 (2007) 30–37.
[117] L. Rossi, E. Bonmassar, I. Faraoni, Modification of miR gene expression pattern
in human colon cancer cells following exposure to 5-fluorouracil in vitro,
Pharmacol. Res. 56 (2007) 248–253.
[118] A.K. Farraj, M.S. Hazari, N. Haykal-Coates, C. Lamb, D.W. Winsett, Y. Ge, A.D.
Ledbetter, A.P. Carll, M. Bruno, A. Ghio, D.L. Costa, ST depression, arrhythmia,
vagal dominance, and reduced cardiac micro-RNA in particulate-exposed rats,
Am. J. Resp. Cell Mol. Biol. 44 (2011) 185–196.
[119] A. Izzotti, R. Balansky, F. D’Agostini, M. Longobardi, C. Cartiglia, S. La Maes-
tra, R.T. Micale, A. Camoirano, G. Ganchev, M. Iltcheva, V.E. Steele, S. De
Flora, Relationships between pulmonary micro-RNA and proteome profiles,
systemic cytogenetic damage and lung tumors in cigarette smoke-exposed
mice treated with chemopreventive agents, Carcinogenesis 34 (2013)
2322–2329.
[120] K. Taki, T. Fukushima, R. Ise, I. Horii, T. Yoshida, Microarray analysis of 6-
mercaptopurine-induced-toxicity-related genes and microRNAs in the rat
placenta, J. Toxicol. Sci. 38 (2013) 159–167.
[121] J. Wu, T. Yang, X. Li, Q. Yang, R. Liu, J. Huang, Y. Li, C. Yang, Y. Jiang, Alter-
ation of serum miR-206 and miR-133b is associated with lung carcinogenesis
induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, Toxicol. Appl.
Pharmacol. 267 (2013) 238–246.
[122] F.M. Cui, J.X. Li, Q. Chen, H.B. Du, S.Y. Zhang, J.H. Nie, J.P. Cao, P.K. Zhou, T.K.
Hei, J. Tong, Radon-induced alterations in micro-RNA expression profiles in
transformed BEAS2B cells, J. Toxicol. Environ. Health 76 (2013) 107–119.
[123] K. Juhasz, K. Gombos, M. Szirmai, K. Gocze, V. Wolher, P. Revesz, I. Magda,
A. Sebestyen, A. Nemeth, I. Ember, Very early effect of DMBA and MNU on
microRNA expression, In vivo 27 (2013) 113–117.
[124] S. Starckx, A. Batheja, G.R. Verheyen, S.D. Jonghe, K. Steemans, B.V. Dijck, M.
Singer, N. Bogdan, J. Snoeys, P. Vinken, J.C. Sasaki, J.V. Gompel, P. Guzzie-
Peck, A. Lampo, L. Lammens, Evaluation of miR-122 and Other Biomarkers in
Distinct Acute Liver Injury in Rats, Toxicol. Pathol. 41 (2013) 795–804.
[125] N.P. Singh, U.P. Singh, H. Guan, P. Nagarkatti, M. Nagarkatti, Prenatal exposure
to TCDD triggers significant modulation of microRNA expression profile in the
thymus that affects consequent gene expression, PLoS One 7 (2012) e45054.
[126] M.J. Jenny, N. Aluru, M.E. Hahn, Effects of short-term exposure to 2,3,7,8-
tetrachlorodibenzo-p-dioxin on microRNA expression in zebrafish embryos,
Toxicol. Appl. Pharmacol. 264 (2012) 262–273.
[127] C. Vacchi-Suzzi, Y. Bauer, B.R. Berridge, S. Bongiovanni, K. Gerrish, H.K.
Hamadeh, M. Letzkus, J. Lyon, J. Moggs, R.S. Paules, F. Pognan, F. Staedtler,
M.P. Vidgeon-Hart, O. Grenet, P. Couttet, Perturbation of micro RNAs in rat
heart during chronic doxorubicin treatment, PLoS One 7 (2012) e40395.
[128] P. Brzuzan, M. Wozny, L. Wolinska, A. Piasecka, Expression profiling in vivo
demonstrates rapid changes in liver microRNA levels of whitefish (Coregonus
lavaretus) following microcystin-LR exposure, Aquat. Toxicol. 122–123 (2012)
188–196.
[129] R.P. Dippold, R. Vadigepalli, G.E. Gonye, J.B. Hoek, Chronic ethanol feeding
enhances miR-21 induction during liver regeneration while inhibiting prolif-
eration in rats, Am. J. Physiol. 303 (2012) G733–G743.
[130] J.A. Bourdon, A.T. Saber, S. Halappanavar, P.A. Jackson, D. Wu, K.S. Hougaard,
N.R. Jacobsen, A. Williams, U. Vogel, H. Wallin, C.L. Yauk, Carbon black
nanoparticle intratracheal installation results in large and sustained changes
in the expression of miR-135b in mouse lung, Environ. Mol. Mutagen. 53
(2012) 462–468.
56 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57
[131] J.C. States, A.V. Singh, T.B. Knudsen, E.C. Rouchka, N.O. Ngalame, G.E. Arteel,
Y. Piao, M.S. Ko, Prenatal arsenic exposure alters gene expression in the adult
liver to a proinflammatory state contributing to accelerated atherosclerosis,
PLoS One 7 (2012) e38713.
[132] J. Ward, S. Bala, J. Petrasek, G. Szabo, Plasma microRNA profiles distinguish
lethal injury in acetaminophen toxicity: a research study, World J. Gastroen-
terol. 18 (2012) 2798–2804.
[133] J. Saikumar, D. Hoffmann, T.M. Kim, V.R. Gonzalez, Q. Zhang, P.L. Goering,
R.P. Brown, V. Bijol, P.J. Park, S.S. Waikar, V.S. Vaidya, Expression, circulation,
and excretion profile of microRNA-21, -155, and -18a following acute kidney
injury, Toxicol. Sci. 129 (2012) 256–267.
[134] M.A. Parasramka, S. Ali, S. Banerjee, T. Deryavoush, F.H. Sarkar, S. Gupta,
Garcinol sensitizes human pancreatic adenocarcinoma cells to gemcitabine
in association with microRNA signatures, Mol. Nutr. Food Res. 57 (2013)
235–248.
[135] F. Wang, W. Liu, J. Ma, M. Yu, Y. Jin, J. Dai, Prenatal and neonatal exposure
to perfluorooctane sulfonic acid results in changes in miRNA expression pro-
files and synapse associated proteins in developing rat brains, Environ. Sci.
Technol. 46 (2012) 6822–6829.
[136] M. Fabbri, C. Urani, M.G. Sacco, C. Procaccianti, L. Gribaldo, Whole genome
analysis and microRNAs regulation in HepG2 cells exposed to cadmium, Altex
29 (2012) 173–182.
[137] G. Zhang, L. Sun, X. Lu, Z. Chen, P.J. Duerksen-Hughes, H. Hu, X. Zhu, J. Yang,
Cisplatin treatment leads to changes in nuclear protein and microRNA expres-
sion, Mutat. Res. 746 (2012) 66–77.
[138] M.K. Song, M. Song, H.S. Choi, Y.J. Kim, Y.K. Park, J.C. Ryu, Identification of
molecular signatures predicting the carcinogenicity of polycyclic aromatic
hydrocarbons (PAHs), Toxicol. Lett. 212 (2012) 18–28.
[139] B.H. Dai, L. Geng, Y. Wang, C.J. Sui, F. Xie, R.X. Shen, W.F. Shen, J.M. Yang,
microRNA-199a-5p protects hepatocytes from bile acid-induced sustained
endoplasmic reticulum stress, Cell Death Dis. 4 (2013) e604.
[140] Y. Teng, T.T. Manavalan, C. Hu, S. Medjakovic, A. Jungbauer, C.M. Klinge,
Endocrine disruptors fludioxonil and fenhexamid stimulate miR-21 expres-
sion in breast cancer cells, Toxicol. Sci. 131 (2013) 71–83.
[141] S.L. Tilghman, M.R. Bratton, H.C. Segar, E.C. Martin, L.V. Rhodes, M. Li, J.A.
McLachlan, T.E. Wiese, K.P. Nephew, M.E. Burow, Endocrine disruptor regu-
lation of microRNA expression in breast carcinoma cells, PLoS One 7 (2012)
e32754.
[142] J.S. Choi, J.H. Oh, H.J. Park, M.S. Choi, S.M. Park, S.J. Kang, M.J. Oh, S.J. Kim, S.Y.
Hwang, S. Yoon, miRNA regulation of cytotoxic effects in mouse Sertoli cells
exposed to nonylphenol, Reprod. Biol. Endocrinol. 9 (2011) 126.
[143] C. Benoit, H. Ould-Hamouda, D. Crepin, A. Gertler, L. Amar, M. Taouis, Early
leptin blockade predisposes fat-fed rats to overweight and modifies hypotha-
lamic microRNAs, J. Endocrinol. 218 (2013) 35–47.
[144] J. Ahn, H. Lee, C.H. Jung, T. Ha, Lycopene inhibits hepatic steatosis via
microRNA-21-induced downregulation of fatty acid-binding protein 7 in mice
fed a high-fat diet, Mol. Nutr. Food Res. 56 (2012) 1665–1674.
[145] Y.M. Yang, S.Y. Seo, T.H. Kim, S.G. Kim, Decrease of microRNA-122 causes
hepatic insulin resistance by inducing protein tyrosine phosphatase 1B, which
is reversed by licorice flavonoid, Hepatology 56 (2012) 2209–2220.
[146] C. Boesch-Saadatmandi, A.E. Wagner, S. Wolffram, G. Rimbach, Effect of
quercetin on inflammatory gene expression in mice liver in vivo - role of
redox factor 1, miRNA-122 and miRNA-125b, Pharmacol. Res. 65 (2012)
523–530.
[147] H. Yin, M. Hu, R. Zhang, Z. Shen, L. Flatow, M. You, MicroRNA-217 promotes
ethanol-induced fat accumulation in hepatocytes by down-regulating SIRT1,
J. Biol. Chem. 287 (2012) 9817–9826.
[148] A.E. Wagner, C. Boesch-Saadatmandi, J. Dose, G. Schultheiss, G. Rimbach, Anti-
inflammatory potential of allyl-isothiocyanate–role of Nrf2, NF-(kappa) B and
microRNA-155, J. Cell. Mol. Med. 16 (2012) 836–843.
[149] Y.J. Kim, S.H. Hwang, H.H. Cho, K.K. Shin, Y.C. Bae, J.S. Jung, MicroRNA 21
regulates the proliferation of human adipose tissue-derived mesenchymal
stem cells and high-fat diet-induced obesity alters microRNA 21 expression
in white adipose tissues, J. Cell. Physiol. 227 (2012) 183–193.
[150] P. Parra, F. Serra, A. Palou, Expression of adipose microRNAs is sensitive to
dietary conjugated linoleic acid treatment in mice, PLoS One 5 (2010) e13005.
[151] R. Takanabe, K. Ono, Y. Abe, T. Takaya, T. Horie, H. Wada, T. Kita, N. Satoh, A.
Shimatsu, K. Hasegawa, Up-regulated expression of microRNA-143 in associa-
tion with obesity in adipose tissue of mice fed high-fat diet, Biochem. Biophys.
Res. Commun. 376 (2008) 728–732.
[152] E.K. Ng, W.W. Chong, H. Jin, E.K. Lam, V.Y. Shin, J. Yu, T.C. Poon, S.S. Ng, J.J.
Sung, Differential expression of microRNAs in plasma of patients with col-
orectal cancer: a potential marker for colorectal cancer screening, Gut 58
(2009) 1375–1381.
[153] T. Ueda, S. Volinia, H. Okumura, M. Shimizu, C. Taccioli, S. Rossi, H. Alder, C.G.
Liu, N. Oue, W. Yasui, K. Yoshida, H. Sasaki, S. Nomura, Y. Seto, M. Kaminishi,
G.A. Calin, C.M. Croce, Relation between microRNA expression and progres-
sion and prognosis of gastric cancer: a microRNA expression analysis, Lancet
Oncol. 11 (2010) 136–146.
[154] J.C. Brase, M. Johannes, T. Schlomm, M. Falth, A. Haese, T. Steuber, T. Beiss-
barth, R. Kuner, H. Sultmann, Circulating miRNAs are correlated with tumor
progression in prostate cancer, Int. J. Cancer 128 (2011) 608–616.
[155] X. Li, Y. Zhang, J. Ding, K. Wu, D. Fan, Survival prediction of gastric cancer by
a seven-microRNA signature, Gut 59 (2010) 579–585.
[156] E. Bandres, E. Cubedo, X. Agirre, R. Malumbres, R. Zarate, N. Ramirez, A.
Abajo, A. Navarro, I. Moreno, M. Monzo, J. Garcia-Foncillas, Identification by
Real-time PCR of 13 mature microRNAs differentially expressed in colorectal
cancer and non-tumoral tissues, Mol. Cancer 5 (2006) 29.
[157] M. Bloomston, W.L. Frankel, F. Petrocca, S. Volinia, H. Alder, J.P. Hagan, C.G. Liu,
D. Bhatt, C. Taccioli, C.M. Croce, MicroRNA expression patterns to differentiate
pancreatic adenocarcinoma from normal pancreas and chronic pancreatitis,
JAMA 297 (2007) 1901–1908.
[158] X.X. Pu, G.L. Huang, H.Q. Guo, C.C. Guo, H. Li, S. Ye, S. Ling, L. Jiang,
Y. Tian, T.Y. Lin, Circulating miR-221 directly amplified from plasma is
a potential diagnostic and prognostic marker of colorectal cancer and
is correlated with p53 expression, J. Gastroenterol. Hepatol. 25 (2010)
1674–1680.
[159] R. Morimura, S. Komatsu, D. Ichikawa, H. Takeshita, M. Tsujiura, H. Nagata,
H. Konishi, A. Shiozaki, H. Ikoma, K. Okamoto, T. Ochiai, H. Taniguchi, E.
Otsuji, Novel diagnostic value of circulating miR-18a in plasma of patients
with pancreatic cancer, Br. J. Cancer 105 (2011) 1733–1740.
[160] H. Cheng, L. Zhang, D.E. Cogdell, H. Zheng, A.J. Schetter, M. Nykter, C.C. Har-
ris, K. Chen, S.R. Hamilton, W. Zhang, Circulating plasma MiR-141 is a novel
biomarker for metastatic colon cancer and predicts poor prognosis, PLoS One
6 (2011) e17745.
[161] G. Gabriely, T. Wurdinger, S. Kesari, C.C. Esau, J. Burchard, P.S. Linsley, A.M.
Krichevsky, MicroRNA 21 promotes glioma invasion by targeting matrix met-
alloproteinase regulators, Mol. Cell. Biol. 28 (2008) 5369–5380.
[162] P.S. Mitchell, R.K. Parkin, E.M. Kroh, B.R. Fritz, S.K. Wyman, E.L. Pogosova-
Agadjanyan, A. Peterson, J. Noteboom, K.C. O’Briant, A. Allen, D.W. Lin, N.
Urban, C.W. Drescher, B.S. Knudsen, D.L. Stirewalt, R. Gentleman, R.L. Ves-
sella, P.S. Nelson, D.B. Martin, M. Tewari, Circulating microRNAs as stable
blood-based markers for cancer detection, Proc. Natl. Acad. Sci. U.S.A. 105
(2008) 10513–10518.
[163] A. Schaefer, M. Jung, H.J. Mollenkopf, I. Wagner, C. Stephan, F. Jentzmik, K.
Miller, M. Lein, G. Kristiansen, K. Jung, Diagnostic and prognostic implica-
tions of microRNA profiling in prostate carcinoma, Int. J. Cancer 126 (2010)
1166–1176.
[164] M. Tanaka, K. Oikawa, M. Takanashi, M. Kudo, J. Ohyashiki, K. Ohyashiki, M.
Kuroda, Down-regulation of miR-92 in human plasma is a novel marker for
acute leukemia patients, PLoS One 4 (2009) e5532.
[165] Z. Huang, D. Huang, S. Ni, Z. Peng, W. Sheng, X. Du, Plasma microRNAs are
promising novel biomarkers for early detection of colorectal cancer, Int. J.
Cancer 127 (2010) 118–126.
[166] R. Mahn, L.C. Heukamp, S. Rogenhofer, A. von Ruecker, S.C. Muller, J. Ellinger,
Circulating microRNAs (miRNA) in serum of patients with prostate cancer,
Urology 77 (2011), 1265 e1269-1216.
[167] M.J. Lodes, M. Caraballo, D. Suciu, S. Munro, A. Kumar, B. Anderson, Detection
of cancer with serum miRNAs on an oligonucleotide microarray, PLoS One 4
(2009) e6229.
[168] H.M. Heneghan, N. Miller, A.J. Lowery, K.J. Sweeney, M.J. Kerin, MicroRNAs as
novel biomarkers for breast cancer, J. Oncol. 2009 (2009) 950201.
[169] N. Yanaihara, N. Caplen, E. Bowman, M. Seike, K. Kumamoto, M. Yi, R.M.
Stephens, A. Okamoto, J. Yokota, T. Tanaka, G.A. Calin, C.G. Liu, C.M. Croce,
C.C. Harris, Unique microRNA molecular profiles in lung cancer diagnosis and
prognosis, Cancer Cell 9 (2006) 189–198.
[170] R. Spizzo, M.S. Nicoloso, L. Lupini, Y. Lu, J. Fogarty, S. Rossi, B. Zagatti, M.
Fabbri, A. Veronese, X. Liu, R. Davuluri, C.M. Croce, G. Mills, M. Negrini, G.A.
Calin, miR-145 participates with TP53 in a death-promoting regulatory loop
and targets estrogen receptor-alpha in human breast cancer cells, Cell Death
Differ. 17 (2010) 246–254.
[171] S.L. Yu, H.Y. Chen, G.C. Chang, C.Y. Chen, H.W. Chen, S. Singh, C.L. Cheng, C.J.
Yu, Y.C. Lee, H.S. Chen, T.J. Su, C.C. Chiang, H.N. Li, Q.S. Hong, H.Y. Su, C.C.
Chen, W.J. Chen, C.C. Liu, W.K. Chan, K.C. Li, J.J. Chen, P.C. Yang, MicroRNA
signature predicts survival and relapse in lung cancer, Cancer Cell 13 (2008)
48–57.
[172] C. Roth, B. Rack, V. Muller, W. Janni, K. Pantel, H. Schwarzenbach, Circulating
microRNAs as blood-based markers for patients with primary and metastatic
breast cancer, Breast Cancer Res. 12 (2010) R90.
[173] M. Garofalo, G. Di Leva, G. Romano, G. Nuovo, S.S. Suh, A. Ngankeu, C. Taccioli,
F. Pichiorri, H. Alder, P. Secchiero, P. Gasparini, A. Gonelli, S. Costinean, M.
Acunzo, G. Condorelli, C.M. Croce, miR-221&222 regulate TRAIL resistance and
enhance tumorigenicity through PTEN and TIMP3 downregulation, Cancer
Cell 16 (2009) 498–509.
[174] H. Si, X. Sun, Y. Chen, Y. Cao, S. Chen, H. Wang, C. Hu, Circulating microRNA-
92a and microRNA-21 as novel minimally invasive biomarkers for primary
breast cancer, J. Cancer Res. Clin. Oncol. 139 (2013) 223–229.
[175] S. Gilad, E. Meiri, Y. Yogev, S. Benjamin, D. Lebanony, N. Yerushalmi, H. Ben-
jamin, M. Kushnir, H. Cholakh, N. Melamed, Z. Bentwich, M. Hod, Y. Goren, A.
Chajut, Serum microRNAs are promising novel biomarkers, PLoS One 3 (2008)
e3148.
[176] J. Silva, V. Garcia, A. Zaballos, M. Provencio, L. Lombardia, L. Almonacid, J.M.
Garcia, G. Dominguez, C. Pena, R. Diaz, M. Herrera, A. Varela, F. Bonilla,
Vesicle-related microRNAs in plasma of nonsmall cell lung cancer patients
and correlation with survival, Eur. Respir. J. 37 (2011) 617–623.
[177] K. Murata, H. Yoshitomi, S. Tanida, M. Ishikawa, K. Nishitani, H. Ito, T.
Nakamura, Plasma and synovial fluid microRNAs as potential biomarkers of
rheumatoid arthritis and osteoarthritis, Arthritis Res. Ther. 12 (2010) R86.
[178] S. Fichtlscherer, S. De Rosa, H. Fox, T. Schwietz, A. Fischer, C. Liebetrau, M.
Weber, C.W. Hamm, T. Roxe, M. Muller-Ardogan, A. Bonauer, A.M. Zeiher, S.
Dimmeler, Circulating microRNAs in patients with coronary artery disease,
Circ. Res. 107 (2010) 677–684.
 B. Siddeek et al. / Mutation Research 764–765 (2014) 46–57
[179] A.M. Zahm, M. Thayu, N.J. Hand, A. Horner, M.B. Leonard, J.R. Friedman,
Circulating microRNA is a biomarker of pediatric Crohn disease, J. Pediatr.
Gastroenterol. Nutr. 53 (2011) 26–33.
[180] D.S. Karolina, A. Armugam, S. Tavintharan, M.T. Wong, S.C. Lim, C.F. Sum, K.
Jeyaseelan, MicroRNA 144 impairs insulin signaling by inhibiting the expres-
sion of insulin receptor substrate 1 in type 2 diabetes mellitus, PLoS One 6
(2011) e22839.
[181] J.P. Cogswell, J. Ward, I.A. Taylor, M. Waters, Y. Shi, B. Cannon, K. Kelnar, J.
Kemppainen, D. Brown, C. Chen, R.K. Prinjha, J.C. Richardson, A.M. Saunders,
A.D. Roses, C.A. Richards, Identification of miRNA changes in Alzheimer’s
disease brain and CSF yields putative biomarkers and insights into disease
pathways, J. Alzheimers Dis. 14 (2008) 27–41.
[182] H. Mizuno, A. Nakamura, Y. Aoki, N. Ito, S. Kishi, K. Yamamoto, M. Sekiguchi,
S. Takeda, K. Hashido, Identification of muscle-specific microRNAs in serum
57
of muscular dystrophy animal models: promising novel blood-based markers
for muscular dystrophy, PLoS One 6 (2011) e18388.
[183] M. Abu-Halima, M. Hammadeh, J. Schmitt, P. Leidinger, A. Keller, E. Meese,
C. Backes, Altered microRNA expression profiles of human spermatozoa in
patients with different spermatogenic impairments, Fertil. Steril. 99 (2013)
1249–1255, e1216.
[184] T. Liu, W. Cheng, Y. Gao, H. Wang, Z. Liu, Microarray analysis of microRNA
expression patterns in the semen of infertile men with semen abnormalities,
Mol. Med. Rep. 6 (2012) 535–542.
[185] W. Wu, Z. Hu, Y. Qin, J. Dong, J. Dai, C. Lu, W. Zhang, H. Shen, Y. Xia, X. Wang,
Seminal plasma microRNAs: potential biomarkers for spermatogenesis sta-
tus, Mol. Hum. Reprod. 18 (2012) 489–497.