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Mutation Research 658 (2008) 215–233

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Review
Genetic polymorphisms and micronucleus formation:
A review of the literature
G. Iarmarcovai a,*, S. Bonassi b, A. Botta a, R.A. Baan c, T. Orsière a
a
Laboratory of Biogenotoxicology and Environmental Mutagenesis (EA 1784; IFR PMSE 112), Faculty of Medecine,
Université de la Méditerranée, 27 Bd Jean Moulin, 13385 Marseille Cedex 5, France
b
Unit of Molecular Epidemiology, National Cancer Research Institute, Genoa, Italy
c
Carcinogen Identification and Evaluation Group, WHO - International Agency for Research on Cancer, Lyon, France
Received 21 June 2007; received in revised form 8 October 2007; accepted 15 October 2007
Available online 22 October 2007

Abstract
The formation of micronuclei (MN) is extensively used in molecular epidemiology as a biomarker of chromosomal
damage, genome instability, and eventually of cancer risk. The occurrence of MN represents an integrated response to
chromosome-instability phenotypes and altered cellular viabilities caused by genetic defects and/or exogenous exposures to
genotoxic agents. The present article reviews human population studies addressing the relationship between genetic
polymorphisms and MN formation, and provides insight into how genetic variants could modulate the effect of environmental
exposures to genotoxic agents, host factors (gender, age), lifestyle characteristics (smoking, alcohol, folate), and diseases
(coronary artery disease, cancer). Seventy-two studies measuring MN frequency either in peripheral blood lymphocytes or
exfoliated cells were retrieved after an extensive search of the MedLine/PubMed database. The effect of genetic poly-
morphisms on MN formation is complex, influenced to a different extent by several polymorphisms of proteins or enzymes
involved in xenobiotic metabolism, DNA repair proteins, and folate-metabolism enzymes. This heterogeneity reflects the
presence of multiple external and internal exposures, and the large number of chromosomal alterations eventually resulting in
MN formation. Polymorphisms of EPHX, GSTT1, and GSTM1 are of special importance in modulating the frequency of
chromosomal damage in individuals exposed to genotoxic agents and in unexposed populations. Variants of ALDH2 genes are
consistently associated with MN formation induced by alcohol drinking. Carriers of BRCA1 and BRCA2 mutations (with or
without breast cancer) show enhanced sensitivity to clastogens. Some evidence further suggests that DNA repair (XRCC1 and
XRCC3) and folate-metabolism genes (MTHFR) also influence MN formation. As some of the findings are based on relatively
small numbers of subjects, larger scale studies are required that include scoring of additional endpoints (e.g., MN in

Abbreviations: MN, micronuclei; PBL, peripheral blood lymphocytes; CA, chromosomal aberrations; SNP, single nucleotide polymorphism;
NQO1, NAD(P)H quinone oxidoreductase; CYP, P450 cytochromes; GST, glutathione S-transferases; NAT, N-acetyl transferases; EPHXgene/mEH
enzyme, microsomal epoxide hydrolase; ADH, alcohol dehydrogenases; ALDH, aldehyde dehydrogenases; MGMT, methylguanine DNA
methyltransferase; DRR, direct reversal repair; BER, base-excision repair; APEX1 or APE1, apurinic/apyrimidinic endonuclease; hOGG1, human
8-oxoguanine glycosylase; XRCC1, X-ray repair cross-complementing protein (group 1); NER, nucleotide-excision repair; XPD/ERCC2,
xeroderma pigmentosum group D/excision repair cross-complementing rodent repair deficiency; DSBR, double-strand-break repair; BRCA1/
BRCA2, breast cancer genes; XRCC3, X-ray repair cross-complementing protein (group 3); MTHFR, methylenetetrahydrofolate reductase; MTR,
methionine synthase; MTRR, methionine synthase reductase; PAH, polycyclic aromatic hydrocarbons; IR, ionizing radiation; ROS, reactive oxygen
species; FISH, fluorescent in situ hybridization.
* Corresponding author. Tel.: +33 4 91 32 45 48; fax: +33 4 91 32 45 72.
E-mail address: Gwenaelle.Iarmarcovai@medecine.univ-mrs.fr (G. Iarmarcovai).

1383-5742/$ – see front matter # 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.mrrev.2007.10.001
216 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233

combination with fluorescent in situ hybridization, analysis of nucleoplasmic bridges and nuclear buds), and address gene–
gene interactions.
# 2007 Elsevier B.V. All rights reserved.

Keywords: Cytokinesis-blocked micronucleus assay; Biological markers; Polymorphism; Molecular epidemiology; Smoking; Exposure

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 216
1.1. General considerations on genetic polymorphisms and cancer risk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 217
1.2. Associations between genetic polymorphisms and micronucleus formation . . . . . . . . . . . . . . . . . . . . . . . 217
2. Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 217
2.1. Search strategy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 217
2.2. Inclusion/exclusion criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219
2.3. Criteria to define associations between genetic polymorphisms and micronucleus formation . . . . . . . . . . . 220
3. Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220
3.1. Genetic polymorphisms and micronucleus formation in association with various environmental
exposures to genotoxic agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..... 220
3.2. Genetic polymorphisms and micronucleus formation in association with host factors, lifestyle
characteristics, and diseases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223
4. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225
4.1. Genetic polymorphisms, micronucleus formation, and environmental exposures . . . . . . . . . . . . . . . . . . . . 225
4.1.1. Environmental exposures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225
4.1.2. Centromere content of micronuclei . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
4.2. Genetic polymorphisms, micronucleus formation, and host factors (age and gender) . . . . . . . . . . . . . . . . 226
4.3. Genetic polymorphisms, micronucleus formation, and lifestyle characteristics . . . . . . . . . . . . . . . . . . . . . 226
4.3.1. Smoking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
4.3.2. Alcohol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
4.3.3. Folate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
4.4. Genetic polymorphisms, micronucleus formation, and diseases (coronary artery disease and cancer) . . . . . 227
5. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228

1. Introduction cancers [3]. Micronuclei are surrogate endpoints in

The formation of micronuclei (MN) is extensively
used in molecular epidemiology as a biomarker of
chromosomal damage and genome instability. The use
of the MN assay in biomonitoring studies has greatly
increased in the last 15 years, and recent international
efforts such as the HUMN (human micronucleus)
project (http://www.humn.org) have greatly contributed
to improving the reliability of this assay, providing
technical guidelines and analyzing major sources of
variability [1,2]. Results from a recent prospective
analysis of data on more than 6700 subjects collected in
20 laboratories representing 10 countries, confirmed
that an elevated MN frequency in peripheral blood
lymphocytes (PBL) is predictive of an increased cancer
risk, in particular urogenital and gastro-intestinal
surrogate cells, presumed to reflect more specific
chromosomal alterations relevant to carcinogenesis
[4,5]. In the prospective analysis mentioned above, the
association between MN and cancer risk could not be
explained by tobacco smoking or occupational exposure
to carcinogens [3]. However, other factors may interact
with cancer predictivity of MN, including other
environmental exposures to carcinogenic agents, diet-
ary intake of micronutrients such as folate, and
individual susceptibility, e.g., through polymorphisms
of genes involved in metabolism of xenobiotics, DNA
repair, and folate metabolism.
The present article reviews and updates published
human population studies on the relationship between
genetic polymorphisms and MN formation. A previous
review by Norppa [5] examined the association of genetic
 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233
polymorphisms and human cytogenetic damage –
chromosomal aberrations (CA), sister chromatid
exchange and MN – in biomonitoring studies performed
either with PBL or with exfoliated cells. However, given
the dramatic increase of biomonitoring studies evaluating
genetic polymorphisms, many relevant papers have been
published since 2004 and the present review considers
nearly 60% more publications in general and nearly twice
as many dealing with MN and polymorphisms in DNA
repair genes. This review also provides insight into how
genetic variants could interact with environmental
exposures to genotoxic agents, host factors (gender,
age), lifestyle characteristics (smoking, diet), and
diseases (cancer, coronary artery disease), and as a
consequence could affect MN frequencies in humans.
1.1. General considerations on genetic
polymorphisms and cancer risk
The most common type of variation in DNA
sequence is the single nucleotide polymorphism
(SNP), present in about 1/1000 nucleotides in the
human genome. The role of genetic variants contribut-
ing to diseases with a complex etiology such as cancer is
poorly known [6]. An essential step in understanding
the complexity of the genotype–phenotype relationship
is to determine which of the many SNPs influence
cancer risk [6]. The left part of Table 1 summarizes
polymorphisms of genes involved in activation/deacti-
vation of xenobiotics, DNA repair, and folate metabo-
lism that have been selected for the review, together
with the conclusions of recent meta-analyses or pooled
analyses of the effects of these genotypic variations on
cancer risk (for more details, see [5,7–36]).
1.2. Associations between genetic polymorphisms
and micronucleus formation
The model linking environmental exposure to
genotoxic agents, lifestyle factors, and genetic back-
ground to the biological/clinical effect is outlined in
Fig. 1. The individual response to stress may vary
according to various conditions, such as the functioning
of the particular gene combination, the absorption and
metabolism rate of genotoxic agents, DNA repair, cell
death (apoptosis/necrosis), cell-cycle control, and
immune response (Fig. 2) [25,37–39]. The formation
of MN in dividing cells is the result of chromosome
breakage due to unrepaired or misrepaired DNA lesions,
or chromosome malsegregation due to mitotic malfunc-
tion. These events may be induced by oxidative stress,
exposure to clastogens or aneugens, genetic defects in
217
cell-cycle checkpoint and/or DNA repair genes, and by
deficiencies in nutrients required as co-factors in DNA
metabolism and chromosome segregation [3]. The MN
assay has in fact evolved into a ‘‘cytome’’ method for
comprehensively measuring chromosome-instability
phenotypes and altered cellular viabilities caused by
genetic defects and/or exogenous exposures to genotoxic
agents and/or nutritional deficiencies, influenced by
susceptibility factors [40–42].
The systematic evaluation of papers reporting on the
association between genotypes and MN is hampered by
common limitations of the study design: (i) the size of
study groups is usually too small to evaluate rare
polymorphisms, and different genotypes are investi-
gated in different studies, (ii) the allele frequencies for
each genotype vary widely in different ethnic popula-
tions, and (iii) statistical analysis is often inappropriate
and potential confounding factors are not fully
evaluated. Pooled analysis, by increasing the statistical
power, is an adequate tool to summarize information
from individual studies, but at the present time, as far as
we know, only one is available [43], and another one in
press [Mateuca et al., in press].1 Despite these
limitations, many papers showed the presence of an
interaction between MN frequencies and certain genetic
polymorphisms involved in metabolic activation/deac-
tivation of xenobiotics, in DNA repair, or in folate
metabolism. The purpose of the present article is to
review the recently published population studies
describing the association between these genetic
polymorphisms and MN formation.
2. Materials and methods
2.1. Search strategy
We performed an extensive literature search without any
language restriction by use of the MedLine/PubMed database
(National Library of Medicine, National Institutes of Health,
Bethesda, MD, USA-http://www.ncbi.nlm.nih.gov/PubMed)
covering the time between 1 January 1985 and 30 September
2007. Table 2 shows the keywords selected from Medical
Subject Headings (MeSH) and the combinations of search
terms used through the Boolean operators. The purpose of this
search was to retrieve from the literature papers reporting
human population studies based on MN analysis and aimed at
1
R.A. Mateuca, M. Roelants, G. Iarmarcovai, P.V. Aka, L. God-
deris, A. Tremp, S. Bonassi, M. Fenech, J.L. Bergé-Lefranc, M.
Kirsch-Volders, hOGG1326, XRCC1399 and XRCC3241 polymorph-
isms influence micronucleus frequencies in human lymphocytes in
vivo, Mutagenesis (2007), doi:10.1093/mutage/gem040.
218 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233

Table 1
List of genetic polymorphisms included in this review (in human peripheral lymphocytes or exfoliated cells)
Gene studied Function Polymorphism Cancer type most frequently Reference of MN population
associated with the gene variant studies included in the review
and evaluating the gene variant
Metabolism
NQO1 Oxidative Pro187Ser Lung, bladder, colorectum [56,64]
scavenger
CYP1A1 Phase 1 m1 (Msp1 site), Lung [50,52,54,56,64,70,85,116–119]
m2 (Ile462Val), and
m4 (Thr461Asn) alleles
CYP2D6 Phase 1 Lung? [51]
CYP2E1 Phase 1 RsaI, PstI or DraI sites – [50,53,55,56,63,64,72,118,120]
GSTA1 Phase 2 Breast [92]
GSTM1 Phase 2 Gene deletion Head and neck, acute leukemia, [43,46–50,52–59,61–65,67,68,
colorectum, lung, larynx, bladder 81–85,92,116–128]
GSTT1 Phase 2 Gene deletion Acute leukemia, breast, [43,47,49,50,52,53,55–65,67,
lung, brain 68,70,81,82,84,85,92,118–122,
124,126,128]
GSTP1 Phase 2 Ile105Val Prostate [50,52,53,55–57,59,61,62,70,
84,92,120,126]
GSTP1 Phase 2 Ala114Val Lung [62]
NAT2 Phase 2 Slow/rapid acetylator Bladder (slow acetylators) [56,57,68,84,85,121]
colon (rapid acetylators)
EPHX/mEH (exon 3)a Phase 2 Tyr113His Risk reduction for lung cancers [52,53,55–57,61–63,70,85,120]
EPHX/mEH (exon 4)a Phase 2 His139Arg Lung [52,53,55–57,61–63,70,85,120]
ADH1B Phase 2 Arg47His Esophagus [18]
ALDH2 Phase 2 Glu487Lys Esophagus [18,71–73]
DNA repair
MGMT DRR Leu84Phe Lung? [69]
APE1 BER Asp148Glu – [129,130]
hOGG1 BER Ser326Cys Lung, prostate, esophagus, [61,63,66,131]
naso-/oro-pharynx
XRCC1 BER Arg194Trp Risk reduction for all cancers [61,63,66,70,129–132]
XRCC1 BER Arg280His All cancers [61,63,66,70,94,130–132]
XRCC1 BER Arg399Gln Lung? [61,63,66,67,69–71,94,119,129–134]
ERCC2/XPD NER Asp312Asn Lung, breast [69,70,133]
ERCC2/XPD NER Lys751Gln Lung, breast [69–71,94,133]
XRCC3 DSBR Thr241Met Breast, bladder, head and neck? [61,63,66,67,94,129–131,133,134]
BRCA1/BRCA2 DSBR Breast, ovaries [30,86–90]
TP53 Cell-cycle Arg72Pro [80]
Folates
MTHFR Folates 677 C > T Lung [35,74–79,91,93]
Risk reduction for colorectal
cancers and acute leukemia
MTHFR Folates 1298 A > C Risk reduction for [74,77,91]
colorectal cancers
MTR Folates 2756 A > G [77,91]
MTRR Folates 66 A > G [74,91,93]
NQO1, NAD(P)H quinone oxidoreductase; CYP, P450 cytochromes; GST, glutathione S-transferases; NAT, N-acetyl transferases; EPHX gene/mEH
enzyme, microsomal epoxide hydrolase; ADH, alcohol dehydrogenases; ALDH, aldehyde dehydrogenases; MGMT, methylguanine DNA
methyltransferase; DRR, direct reversal repair; BER, base-excision repair; APEX1 or APE1, apurinic/apyrimidinic endonuclease; hOGG1, human
8-oxoguanine glycosylase; XRCC1, X-ray repair cross-complementing protein (group 1); NER, nucleotide-excision repair; XPD/ERCC2,
xeroderma pigmentosum group D/excision repair cross-complementing rodent repair deficiency; DSBR, double-strand-break repair; BRCA1/
BRCA2, breast cancer genes; XRCC3, X-ray repair cross-complementing protein (group 3); MTHFR, methylenetetrahydrofolate reductase; MTR,
methionine synthase; MTRR, methionine synthase reductase.
a
A single nucleotide change at codon 113 of exon 3 (leading to a Tyr ! His amino acid change) reduces enzyme activity and a codon 139
substitution (His ! Arg) in exon 4 is associated with increased enzyme activity. On the basis of these exon 3 and exon 4 genotypes, the expected
individual mEH activity can be classified as low, intermediate, or high.
 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233 219

Fig. 1. Relationships among genetic risk factors (genetic polymorphisms), environmental exposures to genotoxic agents, lifestyle characteristics
(smoking, alcohol, micronutrients), and induction of DNA mutations.

assessing the effects of a specific exposure or condition in
relation to genetic polymorphisms. Results of in vitro studies
(such as assays on mutagen sensitivity associated with gene
polymorphisms) were also included; such studies were not
considered in the previous review by Norppa [5]. The NOT
operator was used to exclude most studies on animals or
plants. Tutorial reviews were also excluded.
2.2. Inclusion/exclusion criteria
One hundred twenty-six citations were retrieved through
MedLine search. All the publications specifically addressing
the association between MN formation and genetic poly-
morphisms in humans were manually reviewed. The criteria
for inclusion of studies were as follows: (i) adherence to
standard protocols for MN analysis [44,45], (ii) evaluation
of the effects of an environmental exposure such to geno-
toxic agents, or host-related factors such as age, gender, or
lifestyle characteristics such as smoking, alcohol consump-
tion, diet or the presence of a disease, and (iii) formal
statistical analysis. Only papers providing a clear descrip-
tion of the study design and the study findings were con-
sidered further. Publications reporting results of in vitro
assays on cell lines were excluded.

Fig. 2. Genotoxic agents, DNA damage, and genetic polymorphisms (metabolism and DNA repair) most of the carcinogens present in the
environment undergo bio-activation into ultimately reactive metabolites, and this transformation is modulated by the individual genetic profile of
metabolic genes. Common polymorphisms in DNA repair genes may alter protein function and eventually the individual ability to repair DNA
damage. PAH, polycyclic aromatic hydrocarbons; IR, ionizing radiation; ROS, reactive oxygen species.
220 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233
Table 2
Keywords (MeSH vocabulary) and search terms (including phrase search and free text tools) used in MedLine search
Biomarker-assay Time period
Micronucleus tests [mesh] OR (1989–2007)
‘‘Micronucleus test’’ OR (1985–1988)
‘‘Micronucleus assay’’ OR (1985–1988)
Micronuclei [tiab] (1985–2007)
Tiab, title and abstract; pt, publication type.
2.3. Criteria to define associations between genetic
polymorphisms and micronucleus formation
Of the 126 MedLine retrieved studies, 62 (i.e., about 50%)
met the inclusion criteria and were included in the review. Ten
additional studies were manually identified through the exam-
ination of the references reported by the MedLine-related
links, resulting in a total number of 72 studies (51 biomonitor-
ing studies, 21 in vitro studies) performed with PBL or
exfoliated cells (right part of Table 1). For most studies,
the study design was cross-sectional. Metabolism genes were
the most frequently studied polymorphic genes and were
mentioned in 47 studies (65% of all studies reviewed). GSTM1
was evaluated in 40 studies. Thirteen of the 15 (i.e., 87%)
biomonitoring studies dealing with MN and DNA repair gene
polymorphisms have been published since 2004, the date of
the previous review by Norppa [5].
To simplify the presentation and the interpretation of the
studies reviewed, risk factors were classified according to the
following categories: (i) various environmental exposures to
genotoxic agents (45 studies), (ii) host factors, lifestyle char-
acteristics and diseases (cancer and coronary artery disease)
(25 studies), and (iii) tobacco smoking when studied (14
studies, with three studies specifically addressing the relation-
ship between smoking, genetic polymorphisms, and MN). For
AND operator NOT operator
Polymorphism, genetic [mesh] OR Animals, laboratory
[mesh] OR
Polymorphism [tiab] OR Plants [mesh]
Polymorphisms [tiab] OR
Polymorphic [tiab] OR Review [pt]
SNP [tiab] OR
‘‘Single nucleotide polymorphisms’’ OR
Cytochrome P450 CYP1A1 [mesh] OR
CYP1A1 [tiab] OR
Cytochrome P450 CYP2E1 [mesh] OR
CYP2E1 [tiab]
Glutathione transferase [mesh] OR
Glutathione S-transferase M1 [substance name]
OR GSTM1 [tiab] OR
Glutathione S-transferase T1 [substance name]
OR GSTT1 [tiab] OR
X-ray repair cross-complementing protein 1
[substance name] OR XRCC1 [tiab] OR
X-ray repair cross-complementing protein 3
[substance name] OR XRCC3 [tiab] OR
Oxoguanine glycosylase 1, human
[mesh] OR hOGG1 [tiab] OR
BRCA1 protein [mesh] OR BRCA1 [tiab] OR
BRCA2 protein [mesh] OR BRCA2 [tiab] OR
Methylenetetrahydrofolate reductase
[mesh] OR MTHFR [tiab]
Humans [mesh]
each study, we reported main design features (source of
exposure, size of the exposed and referent groups, and per-
centage of smokers), and main findings (risk genotype asso-
ciated with the exposure studied). Peripheral blood was the
most commonly used biological specimen, with only six
studies evaluating biomarkers in cells from buccal or urothe-
lial mucosa [46–51].
3. Results
3.1. Genetic polymorphisms and micronucleus
formation in association with various environmental
exposures to genotoxic agents
The characteristics and the outcome of the studies by
category of exposure to genotoxic agents are shown in
Table 3. Ten in vitro and 35 biomonitoring studies (20
since 2004, i.e., 57%) were reviewed. Exposure to
organic chemicals was investigated in 51% of all studies.
The most frequently evaluated compounds of this class
were polycyclic aromatic hydrocarbons (PAH) (10
studies), styrene (7), butadiene or its metabolite
(1,2:3,4-diepoxybutane) (3), solvents (toluene and
 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233 221

Table 3
Genetic polymorphisms and micronuclei (in human peripheral blood lymphocytes or exfoliated cells), in association with various environmental
exposures to genotoxic agents
Exposure Number of subjects Cells Polymorphic genes studied Increased MN frequency Reference
studied (exposed/controls) associated with risk genotype
(affected group)a
PAH 71/59 PBL CYP1A1, GSTM1 –b [116]
69/35 PBL CYP1A1, GSTM1 GSTM1-null (chimney sweeps)c [54]
76/18 PBL CYP1A1, GSTM1 – [117]
98/55 PBL CYP1A1, GSTP1, GSTM1, EPHX exon 3 Tyr/Tyr (potroom [52]
GSTT1, EPHX workers)
21/11 Urothelial GSTM1, GSTT1 – [49]
cells
141/66 PBL NQO1, CYP2E1, EPHX exon 3 Tyr/Tyr [56]
GSTP1, GSTM1, (coke-oven workers) and GSTP1
GSTT1, NAT2, EPHX, Val/Val (coke-oven workers)
XRCC1
141/66 PBL XRCC1 – [132]
25/37 PBL GSTM1, GSTT1 – [128]
34/35 PBL XRCC1, APE1, XRCC3 – [130]
166/69 PBL CYP1A1, GSTP1, GSTT1, EPHX exon 3 His/Tyr or Tyr/Tyr [70]
EPHX, XRCC1, ERCC2 (coke-oven workers)d
Styrene 14/30 PBL GSTM1, GSTT1 – [124]
30 healthy donors (in vitro) PBL GSTP1, GSTM1, GSTT1, mEH low activity (induced MN)e [62]
EPHX
44/44 PBL CYP2E1, GSTM1, GSTT1, XRCC1 399 Arg/Gln or [63]
EPHX, hOGG1, XRCC1, Gln/Gln (all)
XRCC3
28/28 PBL CYP2E1, GSTP1, GSTM1, – [120]
GSTT1, EPHX
20 healthy donors (in vitro) PBL GSTP1, GSTM1, GSTT1, GSTT1-positive (induced MN) [61]
EPHX, XRCC1, hOGG1,
XRCC3
32/32 PBL CYP2E1, GSTP1, GSTM1, GSTT1-null (rubber workers) [55]
GSTT1, EPHX
92/77 PBL GSTP1, GSTM1, GSTT1, GSTT1-null (exposed workers) [57]
NAT2, EPHX
Air pollution 134/58 PBL NQO1, CYP1A1, CYP2E1, GSTM1-positive (all) [64]
GSTM1, GSTT1
37 healthy donors Buccal GSTM1, NAT1 NAT1*10 homozygousf [46]
cells
174 healthy donors PBL MTHFR, MTRR MTRR 66 GGf [93]
129 healthy donors (66–63 PBL CYP1A1, GSTM1, – [119]
industrial-rural areas) GSTT1, XRCC1
28–33 (agricultural-industrial PBL GSTM1, GSTT1 GSTT1-null (all) [65]
workers)/44
171 healthy donors PBL XRCC1, ERCC2, XRCC3 – [94]
IR 31/32 PBL hOGG1, XRCC1, XRCC3 hOGG1 326 Ser/Ser (controls), [66]
XRCC3 241 Thr/Met or
Met/Met (all)
21/21 PBL XRCC1, ERCC2, XRCC3 – [133]
64 healthy donors (in vitro) PBL MGMT, ERCC2, XRCC1 XRCC1 399 Arg/Gln or [69]
Gln/Gln, ERCC2 312
Asp/Asp (baseline MN)
20 healthy donors (in vitro) PBL GSTP1, GSTM1, GSTT1, – [61]
EPHX, XRCC1, hOGG1,
XRCC3
12 healthy donors (in vitro) PBL MTHFR MTHFR 677 CT (induced MN) [35]
222 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233
Table 3 (Continued )
Exposure Number of subjects Cells Polymorphic genes studied Increased MN frequency Reference
studied (exposed/controls) associated with risk genotype
(affected group)a
Pesticides 23/22 PBL GSTM1, GSTT1, NAT2 – [121]
30/33 PBL GSTM1, GSTT1, NAT2 GSTM1-positive (all) [68]
64/50 PBL and GSTM1, GSTT1 – [47]
buccal
cells
33/33 PBL CYP2E1, GSTP1, GSTM1, mEH low activity [53]
GSTT1, EPHX (exposed individuals)
Other 53/46 (butadiene) PBL GSTM1, GSTT1 GSTT1-positive (butadiene [58]
exposures producers)
11 GSTM1-positive PBL GSTT1 GSTT1-null and MN/C-MN/C + [60]
(6 GSTT1-positive and 5 MN (induced MN)
GSTT1-null) (1,2:3,
4diepoxybutane, in vitro)
19/19 (butadiene) PBL GSTM1, GSTT1 – [122]
20 healthy donors PBL GSTP1, GSTM1, GSTT1, XRCC3 241 Thr/Met [61]
(ethylene, in vitro) EPHX, XRCC1, hOGG1, or Met/Met (induced MN)
XRCC3
52/36 (solvents, toluene PBL GSTM1 – [125]
and acetone)
39/55 (solvents, toluene) PBL and CYP1A1, CYP2E1, GSTP1, GSTM1-null (controls, MN [50]
buccal GSTM1, GSTT1 in buccal cells)
cells
21–26 (cobalt-hard PBL GSTM1, GSTT1, hOGG1, – [131]
metal exposed)/26 XRCC1, XRCC3
27/30 (welding fumes) PBL XRCC1, XRCC3 – [134]
27/30 (welding fumes) PBL GSTM1, GSTT1, XRCC1, GSTM1-positive and C-MN [67]
XRCC3 (all and controls), GSTT1-null
and C + MN (all)
30/22 (antineoplastic drugs) PBL XRCC1, APE1, XRCC3 – [129]
343/301 (pooled analysis) PBL GSTM1, GSTT1 GSTT1-positive (all) [43]
37 healthy donors PBL GSTM1 – [123]
(benzo(a)pyrene/cumene
hyperoxide, in vitro)
6 healthy donors PBL GSTP1, GSTM1, GSTT1 – [126]
(thimerosal, in vitro)
15 healthy donors PBL GSTP1, GSTM1, GSTT1 GSTM1-null (induced MN) [59]
(hydroquinone, in vitro)
49 patients with systemic lupus Buccal CYP2D6 – [51]
erythematosus/43 healthy donors cells
(cyclophosphamide, in vitro)
a
Results reported from the original publications.
b
Not significant at 0.05 significance level.
c
Biomonitoring studies. Only GSTM1-null workers (noted chimney sweeps) had a higher mean frequency of MN than GSTT1-positive workers.
This association between genetic polymorphisms and MN formation was not reported (in the original publication) within the control group (which
would be noted controls) or within the total population including exposed workers and controls (which would be noted all).
d
Path analysis, an extension of the regression model, was used in this study, and not an univariate statistical analysis.
e
In vitro studies. mEH low activity subjects had increased frequencies of MN after in vitro styrene treatment (noted-induced MN) than mEH
medium activity subjects, and not before in vitro treatment (which would be noted baseline MN).
f
Association with exposure was uncertain, as no controls were examined.

acetone) (2), and ethylene oxide (1). Inorganic chemi-
cals, mostly metals, were studied in three papers. Five
published papers focused on ionizing radiation (IR).
Among exposures most frequently evaluated were air
pollution (6 studies), pesticides (4), and antineoplastic
drugs (1). A pooled analysis of eight studies on the
influence of GSTM1 and GSTT1 [43] polymorphisms on
MN frequencies in human lymphocytes was recently
undertaken within the framework of the HUMN and
CancerRiskBiomarker projects.
 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233
Seven of the 35 biomonitoring studies that were
reviewed reported an increased MN frequency in
combination with various genetic polymorphisms in
exposed individuals only [52–58]. Two studies showed
that the EPHX Tyr113His (exon 3) genotype affected MN
frequencies in PAH-exposed workers [52,56]. Laffon
et al. [55] and Migliore et al. [57] observed elevated MN
frequencies in GSTT1-null workers exposed to styrene.
Six of the 10 in vitro studies (induction of MN in PBL
after various in vitro treatments) included in this part of
the review revealed higher-induced MN frequencies in
GSTM1-null [59], GSTT1-null [60], GSTT1-positive
[61], mEH (low activity) [62], XRCC3-241 wild-type
[61], and MTHFR-677 CT [35] individuals. Another
seven of the 35 biomonitoring studies reported increased
MN frequencies in combination with various genetic
polymorphisms both in exposed individuals and controls
[43,63–68]. Only three biomonitoring studies showed a
significant influence of the genotypes on MN frequencies
in the control populations [50,66,67]. The in vitro study
of Rzeszowska-Wolny et al. [69] showed that the baseline
frequency of micronucleated cells was significantly
lower in individuals with ERCC2-312 wild-type and
higher in individuals with XRCC1-399 wild-type. Qiu
et al. [70] aimed at identifying causal interrelationship of
various biomarkers using the path analysis. Their results
indicated that exogenous agents, especially the coke-
oven exposure, played a more important role than the
genotypes in the induction of early genetic damage.
Polymorphisms in GSTM1 and GSTT1 were widely
studied in relation to exposure to genotoxic agents, but the
results were inconsistent and conflicting. Some studies on
GSTM1 found higher MN frequencies in positive
individuals compared to their null counterparts [64,68],
while others found the opposite effect [50,54,59] within
exposed individuals, controls, or total population. In a
number of studies GSTT1-null subjects had higher MN
frequencies than their positive counterparts [55,57
,60,65], while in other studies higher MN frequencies
were reported for GSTT1-positive individuals [43,58,61].
Only three of the 35 biomonitoring studies combined
the analysis of genetic polymorphism and the detection
of centromeres in MN using fluorescent in situ
hybridization (FISH) with pancentromeric DNA probes
[57,60,67].
3.2. Genetic polymorphisms and micronucleus
formation in association with host factors, lifestyle
characteristics, and diseases
Studies on the effects of genetic polymorphisms on
MN formation in connection with host factors (age),
223
lifestyle characteristics (alcohol consumption and folate
deficiency), and diseases (cancer and coronary artery
disease) are listed in Table 4. Twelve in vitro and 13
biomonitoring studies – six of these published in or after
since 2004, i.e., 46% – were reviewed.
A recent pooled analysis by Kirsch-Volders et al.
[43] indicated that the GSTT1 genotype influenced MN
frequencies in an age-dependent way. Lower MN
frequencies were found in GSTT1-null subjects aged 20,
and higher MN frequencies in GSTT1-null subjects aged
60, when compared with GSTT1-positive genotypes.
The age-dependent influence of the GSTT1 genotype on
MN frequencies was found only for the group
occupationally exposed. Another important finding of
this pooled analysis [43] was that women occupation-
ally exposed to genotoxic agents were at higher risk for
MN induction than men.
Higher MN frequencies were consistently reported
for ALDH2-deficient habitual drinkers [18,71–73].
Some studies on MTHFR 677 found higher MN
frequencies in individuals carrying the variant TT
genotype as compared with (i) CC or CT genotypes
[74,75] or (ii) CC genotype only [76], while others
showed that the MTHFR 677 C > T polymorphism did
not influence the MN frequency [77–79]. Leopardi et al.
[35] reported that folic acid deficiency significantly
affected the frequency of MN induced by IR. This
increased susceptibility to IR was accounted for by the
stronger response observed in MTHFR 677 C > T cells
compared with CC wild-type cells. Crott et al. [78]
explained the lack of difference between CC and TT
individuals by the fact that a high concentration of
riboflavin (acting as a confounding factor) improves
MTHFR activity and abolishes the protection afforded
by the C677T polymorphism.
Six studies focused on coronary artery diseases in
relation with genetic polymorphisms [74,75,79–82].
Two out of three studies on MTHFR 677 observed
higher MN frequencies in individuals carrying the
variant TT genotype as compared with CC or CT
genotypes [74,75]. Only three studies investigated the
influence of genotypes on the baseline MN level in
cancer patients (Table 4) [83–85]. Six studies dealt
with the induction of MN by IR [86–89], hydrogen
peroxide [86], antineoplastic drugs [90], and folate
deficiency [30] in lymphocytes of women from
families with familial breast cancer and with muta-
tions in the breast cancer susceptibility genes BRCA1
or BRCA2. Three out of six studies observed an
increase in chromosomal mutagen sensitivity in
patients heterozygous for BRCA1 or BRCA2 mutations
[86,87,90]. No enhanced sensitivity towards aneugens
224 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233

Table 4
Genetic polymorphisms and micronuclei (in human peripheral blood lymphocytes or exfoliated cells), in association with host factors, lifestyle
characteristics (alcohol, folic acid), and diseases (coronary artery disease, cancer)
Host factors, lifestyle, Number of subjects Cells Polymorphic genes Increased MN Reference
and diseases (exposed/controls) studied frequency with risk
genotype (affected group)
Host factors 90 healthy donors PBL CYP1A1, CYP2E1, CYP2E1*3 variant [118]
60 healthy donors GSTM1, GSTT1
(30 young-30 elderly, in vitro) PBL GSTM1 – [127]

Lifestyle Alcohol 42 healthy donors PBL ALDH2, XRCC1, ALDH2-deficient [71]

(18 habitual drinkers) ERCC2 (habitual drinkers)
47 healthy donors PBL ALDH2 ALDH2-deficient [73]
(acetaldehyde, in vitro) (induced MN)
248 healthy donors PBL CYP2E1, ALDH2 ALDH2-deficient [72]
(116 habitual drinkers) (habitual drinkers),
wild-type CYP2E1*1/*1
(habitual drinkers)
286 healthy donors PBL ADH1B, ALDH2 ALDH2-deficient [18]
(140 habitual drinkers) (habitual drinkers)
Folic acid 52 healthy donors PBL MTHFR, MTR – [77]
(in vitro)
10(CC)/10(TT) healthy donors PBL MTHFR – [78]
7(CC)/7(TT) healthy donors PBL MTHFR MTHFR 677 [76]
TT (induced MN)
BRCA1 12 mutation carriers with PBL BRCA1, BRCA2 – [30]
breast cancers/8 without/15
healthy donors (BRCA2 10/12/9)
12 healthy donors PBL MTHFR MTHFR 677 [35]
CT (induced MN)

Diseases Coronary 60/22 PBL TP53 – [80]

artery
disease
51/17 PBL MTHFR, MTRR MTHFR 677 TT (all) [74]
63 PBL MTHFR MTHFR 677 TT [75]
46/60 PBL MTHFR – [79]
66 PBL GSTM1, GSTT1 GSTM1-null [81]
39/67 PBL GSTM1, GSTT1 – [82]
Cancer 42/55 (lung cancer) PBL GSTM1 – [83]
28 (mesothelioma) PBL CYP1A1, GSTM1, – [85]
GSTT1, NAT2, EPHX
30 (thyroid cancer, radioiodine PBL GSTP1, GSTM1, NAT2 rapid [84]
treatment) GSTT1, NAT2 (induced MN)
22 women with breast family PBL BRCA1 BRCA1 mutations [86]
history (12 mutation carriers-10 (induced MN)
non-carriers)/17 healthy donors
(IR and hydrogen peroxide, in vitro)
19 women with breast family PBL BRCA1, BRCA2 BRCA1, BRCA2 [87]
history (10 BRCA1 mutation mutations (induced MN)
carriers-9 BRCA2)/14 healthy
donors (IR, in vitro)
13 BRCA1 mutation carriers/13 healthy PBL BRCA1 BRCA1 mutations [90]
donors (antineoplastic drugs, in vitro) (induced MN)
20 women with breast family PBL BRCA1, BRCA2 – [88]
history (11 BRCA1-9 BRCA2
mutation carriers)/78 women with
breast cancers (without mutations)/58
healthy donors (IR, in vitro)
25 cancer free women with PBL BRCA1 – [89]
BRCA1 mutations/25 healthy
donors (IR, in vitro)
For details, see caption of Table 3.
 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233 225

Table 5
Genetic polymorphisms and micronuclei (in human peripheral blood lymphocytes or exfoliated cells), in response to tobacco smoking (in
biomonitoring studies, when assessed)
Biomonitoring studies Polymorphic Increased MN frequency Reference
genes studied associated with risk
Study groups Number Number of genotype (in smokers)
of cases controls
(% smokers) (% smokers)
Floriculturists (pesticides) 30 (29) 33 (30) GSTM1, GSTT1, NAT2 NAT2 rapid [68]
Coke-oven workers (PAH) 21 (38) 11 (45) GSTM1, GSTT1 – [49]
Nuclear reactor cleaners (IR) 30 (73) 30 (20) hOGG1, XRCC1, XRCC3 – [66]
Styrene-exposed workers 44 (48) 44 (48) CYP2E1, GSTM1, – [63]
GSTT1, EPHX, hOGG1,
XRCC1, XRCC3
Cobalt workers 21 (NA) 26 (NA) GSTM1, GSTT1, hOGG1, – [131]
XRCC1, XRCC3
Hard metal workers 26 (NA)
Exposed workers (pesticides) 33 (30) 33 (33) CYP2E1, GSTP1, – [53]
GSTM1, GSTT1, EPHX
Welders 27 (37) 30 (53) GSTM1, GSTT1, GSTM1-positive and [67]
XRCC1, XRCC3 C-MN, GSTT1-null
and MN/C + MN
Healthy donors 154 (100) NA GSTM1 – [48]
(smoking, buccal cells)
Healthy donors (smoking, PBL) 132 (45) NA MTHFR, MTR, MTRR MTRR 66 AA [91]
Healthy donors (smoking, PBL) 72 (35) NA GSTA1, GSTP1, GSTM1-null [92]
GSTM1, GSTT1
Healthy donors (alcohol) 248 (47) NA CYP2E1, ALDH2 – [72]
Healthy donors (alcohol) 286 (51) NA ADH1B, ALDH2 – [18]
Coronary artery disease patients 66 (20) NA GSTM1, GSTT1 GSTM1-null [81]
Lung cancer patients 42(33) 55(31) GSTM1 – [83]
NA, not assessed in the original publications. For details, see caption of Table 3.

(taxol) was measured in lymphocytes of patients with
a BRCA1 mutation [90].
Only 14 of the 51 biomonitoring studies investigated
the relationship between genetic polymorphisms and
MN formation in response to tobacco smoking
(Table 5). Only five studies showed a significant
influence of the genotypes on MN frequencies in
smokers [67,68,81,91,92].
4. Discussion
4.1. Genetic polymorphisms, micronucleus
formation, and environmental exposures
4.1.1. Environmental exposures
Seven of the 35 biomonitoring studies that were
reviewed suggested that genetic polymorphisms may
affect the level of chromosome damage associated with
environmental exposures to genotoxic agents [52–58].
The genetic polymorphisms potentially important for
MN formation depend on the exposure, biological
material examined, and ethnicity of the population
studied. Given the large heterogeneity of individual
exposure to genotoxic agents, reliable estimates of
exposure levels are crucial for the correct interpretation
of genotype–exposure interactions [5]. MN formation
may also reflect genotoxic exposures that occurred
months before the cell sampling. Within cross-sectional
studies, it is possible (i) to collect detailed and accurate
information on current exposure patterns, potential
confounders, and effect modifiers, and (ii) to process
samples for MN analyses within a relatively short
period of time after collection, thereby increasing cell
viability and decreasing the potential for measurement
errors [6]. However, a comparison group (unexposed
control group) is always necessary in order to make
inferences about exposure–genotype interactions. A
genotype effect in the exposed group that is not seen in
the controls suggests exposure-related genotype effects.
Similar effects exposed and control groups or in the
control group only are in keeping with a pure genotype
effect on MN formation [5]. Only three of the 35
biomonitoring studies that were reviewed did not
include a referent control group [46,93,94].
226 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233
Results are inconsistent within exposed population
subgroups, and vary depending on whether the genes
under study are involved in the activation/deactivation
of the agent or in DNA repair. Although mEH (EPHX
gene) is considered a detoxifying enzyme, the
dihydrodiols derived from PAHs may be further
transformed by specific CYP enzymes into still more
reactive species, such as dihydrodiol epoxides, the
ultimate mutagenic and carcinogenic metabolites of the
PAH. GSTT1 detoxifies monohalomethanes (e.g.,
methyl bromide) and the epoxides of the alkenes
ethylene and butadiene, but it activates methylene
chloride and certain bifunctional alkylating agents [8].
As this enzyme has both detoxifying and activating
properties with respect to many environmental pollu-
tants, it is difficult to predict the biological con-
sequences of the GSTT1-null genotype. Parl [95] noted
a higher relative risk for breast cancer in Caucasian
women with GSTM1-positive genotype than in women
with the null genotype. To explain this result, he
suggested that the combined conjugation activities of all
GSTs may lead to depletion of glutathione and thereby
become counterproductive.
Wacholder et al. emphasized the risks of over-
interpretation of statistically significant results, as well
as the need to improve the plausibility of findings from
high-throughput studies defining prior probability that
the observed association between genetic variants and
disease is real [6,96]. A positive interaction between an
exposure to genotoxic agents and genotypes may be
informative, but a null association does not rule out this
interaction as that exposure may act through a
mechanism not reflected by induction of MN [6].
Moreover, a recent pooled analysis [Mateuca et al., in
press] (see footnote 1) indicated that single DNA repair
gene polymorphisms were not likely to have a major
impact on MN frequencies, but rather combinations of
different DNA repair genes.
4.1.2. Centromere content of micronuclei
Chromosome breakage and chromosome loss
involve different cellular and/or molecular dysfunc-
tions. Because acentric chromosomal fragments result-
ing from double-strand breaks can form MN, the
proportion of MN harboring acentric fragments is
expected to be influenced by genetic polymorphisms
involved in the repair of DNA strand breaks. The
proportion of MN harboring whole chromosomes could
also be expected to be affected by the lack of essential
cofactors (e.g., magnesium and calcium) required for
kinetochore and spindle assembly and by genetic
polymorphisms of enzymes controlling the reactivity of
aneugens (e.g., inhibitors of tubulins, topoisomerases
and cyclins) or the activity of cell-cycle check points
(such as silencing hCDC4, a gene involved in cell-cycle
check points, which results in an excessive cyclin E
expression and an increased MN frequency), although
the presence and importance of such polymorphisms are
still unexplored issues [97]. Recent studies also
suggested an additional contribution of the XRCC3
Thr241Met variant to the induction of MN arising from
chromosome loss via malsegregation events and
centrosome amplification [98].
4.2. Genetic polymorphisms, micronucleus
formation, and host factors (age and gender)
An increase in MN frequencies with age and gender
was observed by the pooled analysis of Kirsch-Volders
et al. [43]. Imperfections in cellular defense systems
that protect against the fixation of DNA damage – such
as ROS-induced DNA damage – and decrease efficacy
in DNA repair can lead to an accumulation of mutations
that, on their own or in combination with other age-
related changes, may contribute to ageing and the
development of age-related pathologies [99,100].
Bastaki et al. [101] found suggestive evidence for an
interaction between gender and genotype, with gender
modifying the effect of genotype (Pro197Leu poly-
morphism) on glutathione peroxidase-1 enzyme activ-
ity. This gender-specific difference could be due to
dietary antioxidant intake of selenium [101]. Bond and
Levine [102] showed that the oncogene MDM2-309
T > G (a polymorphism in the p53 tumor suppressor
pathway) accelerated tumor formation in a gender-
specific and hormone-dependent manner (estrogen-
signaling pathway).
4.3. Genetic polymorphisms, micronucleus
formation, and lifestyle characteristics
4.3.1. Smoking
Studies on the relationship between genetic poly-
morphisms and MN formation in response to tobacco
smoking were too sparse to provide conclusive results.
Tobacco smoke contains a mixture of genotoxic
carcinogens that are metabolically activated (phase 1,
especially CYP1A1) or detoxified (phase 2, such as
GST). Polymorphisms of enzymes catalyzing these
reactions are expected to influence the genotoxic and
carcinogenic effects associated with tobacco smoking,
as are polymorphisms of proteins involved in repair of
DNA damage induced by tobacco carcinogens [5].
Husgafvel-Pursiainen [103] indicated associations
 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233
between CYP1A1-Ile462Val polymorphism (exon 7) or
GSTM1-null genotype and lung cancer in never-
smokers and in women. The Human MicroNucleus
project has shown that smokers do not experience an
overall increase in MN frequency, although when the
interaction with occupational exposure is taken into
account, heavy smokers were the only group showing a
significant increase in genotoxic damage as measured
by the MN assay in PBL [104]. The number of heavy
smokers in these studies was in most cases too small to
make an evaluation of the interaction between smoking,
MN, and genetic polymorphisms.
4.3.2. Alcohol
Variants of ALDH2 genes are significantly associated
with genotoxicity/MN formation induced by alcohol
drinking [18,71–73]. In humans, more than 90% of
ingested alcohol is eliminated via metabolic degrada-
tion mainly in the liver. The efficiency in converting
ethanol to acetaldehyde, and subsequent conversion to
acetate, is largely determined by the ADH and ALDH
gene families, with large potential interindividual
differences in acetaldehyde exposure due to the
presence of some well-studied common genetic variants
with a functional role [18]. The variant allele ALDH2*2,
which encodes an inactive sub-unit of the enzyme
ALDH2, is dominant and highly prevalent among
certain populations of Asian ethnicity (0.28–0.45), but
rare in other ethnic groups [19]. These results suggest
that carcinogenic, mutagenic, and genotoxic effects of
ethanol per se on humans need to be further tested,
taking ALDH2 polymorphisms into account.
4.3.3. Folate
Polymorphisms affecting folate-metabolism genes
may modulate genome stability through an effect on
nucleotide pools and DNA methylation (for more
details, see [5]). Under conditions of folic acid
deficiency the MTHFR variant genotype is associated
with an increased risk for developmental defects in
utero and cancer at various sites [35,36]. Several studies
on folic acid deficiency in cultured human lymphocytes
quantified the interrelationship between MN (a bio-
marker of chromosome breakage and/or whole chro-
mosome loss), nucleoplasmic bridges (a biomarker of
DNA misrepair and/or telomere end-fusions), and
nuclear buds (a biomarker of elimination of amplified
DNA and/or DNA repair complexes) in an attempt to
validate the use of these biomarkers and to determine
more comprehensively the impact of folic acid
deficiency, within the physiological range, on various
aspects of genomic stability [42,105].
227
4.4. Genetic polymorphisms, micronucleus
formation, and diseases (coronary artery disease
and cancer)
Coronary artery disease and cancer are the leading
causes of morbidity and mortality in the western world.
Both diseases share a large number of common risk
factors, including cigarette smoking and diet, as well as
common pathogenic processes, such as chronic
inflammation, oxidative stress and genetic instability
[106]. Nutritional deficiencies, MTHFR variants, and
other polymorphisms of the genes involved in folate
metabolism may be important determinants of coronary
artery diseases [74,75]. Gene–diet interaction modu-
lated by genetic polymorphisms of metabolism and
DNA repair and response to micronutrient deficiency
may be equally important in determining genomic
stability and disease risk [107].
Increased sensitivity to mutagens was detected in
carriers of BRCA1/2 mutations, not only after irradia-
tion but also after in vitro treatment with chemical
mutagens (i.e., hydrogen peroxide, antineoplastic
drugs). This enhanced induction of MN suggests that
different DNA repair pathways are affected by BRCA1/
2 mutations, in accordance with the proposed central
role of these genes in maintaining genomic integrity
[32].
An association between MN frequency and cancer
risk was inferred from mechanistic similarities with CA,
which were shown to be predictive for cancer
[3,108,109]. A major challenge in the mechanistic
interpretation of the association between MN formation
and cancer risk is that MN can be generated through
different processes, i.e., chromosome breakage and
chromosome loss (aneuploidy), occurring roughly in
the same proportion [43]. In contrast to chromosome
breakage, whose role in early stages of carcinogenesis
has been extensively studied, the significance of
aneuploidy is still poorly understood, although it is
known that aneuploidy is a hallmark of the majority of
human tumors and associated with high-grade inva-
siveness and poor prognosis [110]. Aneuploidy can
initially occur by a variety of mechanisms: (i) abnormal
centrosome number, leading to multipolar mitoses, (ii)
chromosome loss at anaphase as a result of kinetochore
defects, (iii) malsegregation of chromosomes at
anaphase as a result of defects in the separation of
chromatids, (iv) mitotic slippage caused by inhibition of
mitosis, leading to the formation of tetraploid cells, and
(v) failure of cytokinesis following nuclear replication
as a result of defects in microfilament assembly [111].
Tutt et al. [112] reported that the absence of wild-type
228 G. Iarmarcovai et al. / Mutation Research 658 (2008) 215–233
BRCA2 leads to the development of spontaneous MN
and to the presence of amplified centrosomes.
Furthermore, previous studies reported a link between
numerical homeostasis of centrosomes and genes
involved in DSBR, NER, cell-cycle checkpoints, and
apoptosis [113].
5. Conclusion
The effect of genetic polymorphisms on MN
formation is complex, influenced to a different extent
by several polymorphisms of proteins and enzymes
involved in xenobiotic metabolism, DNA repair
proteins, and folate-metabolism enzymes. This hetero-
geneity reflects the presence of multiple external and
internal exposures, and the large number of chromo-
somal alterations eventually resulting in MN formation.
Polymorphisms of EPHX, GSTT1, and GSTM1 are of
special importance in modulating the frequency of
chromosomal damage either in individuals exposed to
genotoxic agents (exposure-related genotype effects) or
both in exposed and unexposed populations (pure
genotype effects). Variants of ALDH2 genes are
consistently associated with MN formation induced
by alcohol drinking. Carriers of BRCA1 and BRCA2
mutations (with or without breast cancer) show
enhanced sensitivity to clastogens, compared with
healthy donors. Some evidence further suggests that
DNA repair (XRCC1 and XRCC3) and folate-metabo-
lism genes (MTHFR) also influence MN formation
[114]. As some of the findings are based on relatively
small numbers of subjects and might be due to chance,
further studies (pooled analyses) are needed to confirm
these results and to explain the putative influence of
DNA repair gene polymorphisms on chromosomal
damage.
Micronuclei are generally used as a biomarker of
chromosomal instability, integrating acquired muta-
tions and genetic susceptibility, and therefore it remains
to be examined whether these or other genetic
polymorphisms could explain the cancer risk associated
with high-MN frequencies. A better understanding of
MN induction driven by genetic polymorphisms (e.g.,
DNA repair and/or folate-metabolism genes) and
larger-scale studies are required, which should also
include scoring of additional endpoints (e.g., MN-FISH
with pancentromeric DNA probes, nucleoplasmic
bridges, and nuclear buds). It would be also important
to incorporate host factors such as gender, age, and
menopausal status at diagnosis, as well as lifestyle
characteristics, e.g., smoking status – with special
attention to heavy smokers – , alcohol consumption,
exogenous estrogen intake, diet, and micronutrient
status.
Many variant alleles (with meaningful frequency)
are known for each of the major pathways of DNA-
damage response, distributed among the various genes
involved, with the consequence that each individual is
expected to carry about one dozen variants in each
pathway [7]. This means that any major impact on the
overall efficiency of the pathway is likely to arise from a
combination of variants, whereas a single polymorph-
ism would cause only undetectable changes. Selecting
the right combination of variants constitutes a major and
widely discussed challenge in the field of genetic
epidemiology [7]. Thus, when the effect of a single
polymorphism is absent or not strong enough, the
identification and characterization of susceptibility
genes for cancer risk require the understanding of
gene–gene interactions [115].
Acknowledgements
We thank Donatella Ugolini (National Cancer
Research Institute, Genoa, Italy) for her help in search
of the MedLine/PubMed database. The work of Stefano
Bonassi was supported by grants funded by the
Associazione Italiana per la Ricerca sul Cancro (AIRC)
and the Italian Space Agency (ASI).
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