Professional Documents
Culture Documents
research focus
Michael Fenech
Cooperative Research Centre
for Diagnostics
CSIRO Health Sciences and
Nutrition
PO Box 10041
Adelaide BC, SA 5000
Australia
tel: +618 8303 8880
fax: +618 8303 8899
e-mail:
michael.fenech@hsn.csiro.au
1128
www.drugdiscoverytoday.com
1359-6446/02/$ see front matter 2002 Elsevier Science Ltd. All rights reserved. PII: S1359-6446(02)02502-3
research focus
reviews
Table 1. Examples of aneuploidy, chromosome rearrangement, chromosome deletions and gene amplification in
specific cancers (Refs [5666]).
a
Tumour type
Aneuploidy
Chromosome rearrangementa
Lung cancer
Ch 7
3q13.2
3p1323
Breast cancer
Ch 1 and 17
1q2123
1p1113
Prostate cancer
7q22
10q22
Colorectal
carcinoma
Ch 17
17p
17p
Skin melanoma
Ch 7 and 10
1q11q12
1p1122
11q13, 22q1113
Breaks/dicentrics
Malsegregation
BFB cycles
Aneuploidy cycles
Viable
Malignant
Cancer
Drug Discovery Today
www.drugdiscoverytoday.com
1129
reviews
research focus
Genotoxin
or folate
deficiency
Bridge
Breakage
BFB and GA
cycle
Elimination
of amplified
genes by
nuclear budding
Excision of
amplified genes
Fusion &
gene
amplification
Drug Discovery Today
1130
www.drugdiscoverytoday.com
research focus
reviews
www.drugdiscoverytoday.com
1131
reviews
research focus
(a)
Apoptosis
Genotoxic
event
G0
G1,S,G2
35
Necrosis
(b)
ANOVA P<0.0001
30
**
25
20
15
10
5
0
Chromosome
Centrosome
NonDicentric breakage
Gene
abnormality
disjunction*
chromosome and
amplification
loss
Micronucleated cells
*
12
24
60
120
Folic acid concentration (nM)
Nuclear buds
12.5
ANOVA P<0.0001
common mechanism initiated by folic acid deficiencyinduced DNA breaks, although coincidence of effects with
other DNA damage events (e.g. folic acid deficiency-induced
CpG hypomethylation) cannot be excluded.
10.0
**
7.5
5.0
2.5
0.0
12
(c)
24
60
120
Folic acid concentration (nM)
Nucleoplasmic bridges
10.0
ANOVA P<0.0001
*
NPBs per 1000 BNs
Figure 4. The various possible fates of cultured cytokinesisblocked cells following exposure to cytotoxic and/or genotoxic
agents. Using these biomarkers within the cytokinesis-block
micronucleus (CBMN) assay it is possible to measure the
frequency of chromosome breakage (MN), chromosome loss
(MN), chromosome rearrangement, for example, dicentric
chromosomes (NPB), gene amplification (nuclear buds),
necrosis and apoptosis. In addition, cytostatic effects are readily
estimated from the ratio of mono-, bi- and multinucleated cells.
Chromosome loss can be distinguished from chromosome
breakage using pancentromeric probes or anti-kinetochore
antibodies. *In addition, non-disjunction (malsegregation of
chromosomes) can also be measured in BN cells using
chromosome-specific centromeric probes. The CBMN assay has
not been previously used to measure centrosome abnormalities
but this is theoretically possible by scoring the frequency of
tri- and/or quadrinucleated cells and confirming that their DNA
content is 4N. Reproduced, with permission, from Ref [66].
7.5
**
5.0
2.5
0.0
12
24
60
120
Folic acid concentration (nM)
Drug Discovery Today
1132
www.drugdiscoverytoday.com
research focus
NuclearBuds
NPBs
MNed BNs
[Folic acid]
MNed BNs
NPBs
P
r
P
r
<0.001
0.68
<0.001
0.63
<0.001
0.76
<0.001
0.77
<0.001
0.75
<0.001
0.74
reviews
www.drugdiscoverytoday.com
1133
reviews
research focus
1134
www.drugdiscoverytoday.com
research focus
reviews
Genediet
interaction
Micronutrient
deficiency
Increased
infertility?
DNA repair
DNA
damage
Higher cancer
risk
Genetoxin
interaction
Genotoxin
Accelerated
ageing?
Drug Discovery Today
they also showed that MN frequency was not only significantly and negatively correlated with serum vitamin B12
but also significantly and positively correlated with plasma
homocysteine, a risk factor for cardiovascular disease [52,53].
These cross-sectional data showed that MN frequency was
minimized when plasma homocysteine was <7.5 mol l1
and serum B12 was >300 pmol l1, the latter being double
the concentration associated with anaemia. Using a placebocontrolled intervention trial, we also demonstrated that
supplementation over three months with 3.5-times the
Australian recommended dietary allowance (RDA) for folic
acid (700 g) and vitamin B12 (7 g) reduced MN frequency in lymphocytes by 25% in young adults who were
in the top 50% of MN frequency but had no influence on
MN frequency in those in the low 50% of MN frequency
before the intervention [53]. These results illustrate that
dietary deficiency of folate and vitamin B12 is an important determinant of DNA damage rate and that above
RDA intakes of these vitamins might be required in large
subsets of a population to minimize DNA damage rate.
These results have an added important implication
in that it is now feasible and plausible to consider the
concept of recommended dietary allowances (RDAs) using
genomic stability as the main parameter on which to set
these intake levels [54]. The importance of this approach
is supported by the fact that: (a) several micronutrients,
such as zinc, magnesium, folic acid and vitamin B12, are
required as co-factors in DNA metabolism, (b) deficiency
in these co-factors could induce important chromosomal
mutations that increase cancer risk, and (c) genomic stability is crucial for normal function of cells. We can envisage
a future when the ability to use DNA damage biomarkers
for predicting cancer risk will be considerably improved
because of a better understanding of how dietary factors
modulate genomic stability.
www.drugdiscoverytoday.com
1135
reviews
research focus
The future
The future for the use of DNA damage biomarkers will consist of a better integration of classical cytogenetic and noncytogenetic methods with molecular probes, leading to
automated simultaneous and multiple measurements, not
only of events that lead to a hypermutable state but also of
specific mutations on the causal pathway of cancer. The
use of cDNA arrays and proteomics will enable us to better
understand which genes are upregulated and downregulated following specific genotoxic insults and which of
these correlate best with DNA damage events on the causal
pathway of cancer. The main challenge is to make the full
loop in predicting cell phenotype (normal or cancer) from
the large amount of information generated from array
strategies. Equally important will probably be the shift
from considering not only genetoxin interaction in predicting DNA damage and cancer outcomes but to integrate
this approach with genediet interaction studies (Fig. 6).
This comprehensive approach, combined with more robust techniques, will hopefully be fruitful in identifying
better not only the cancer risk of large genetic sub-groups
but also individuals, which is the ultimate goal.
References
1 Solomon, E. et al. (1991) Chromosome aberrations and cancer. Science
254, 11531160
2 Vogelstein, B. and Kinzler, K.W. (1998) in The Genetic Basis of Human
Cancer (Kinzler, K.W. and Vogelstein, B. eds.), pp. xvxiv, McGraw-Hill
3 Cahill, D.P. et al. (1999) Genetic instability and Darwinian selection in
tumours. Trends Cell Biol. 9, M57M60
4 Gray, J.W. and Collins, C. (2000) Genome changes and gene expression
in human solid tumours. Carcinogenesis 12, 443452
5 Loeb, L.A. (1991) Mutator phenotpye may be required for multi-stage
carcinogenesis. Cancer Res. 51, 30753079
6 Li, R.H. et al. (2000) Aneuploidy vs gene mutation hypothesis of cancer:
recent study claims mutation but is found to support aneuploidy.
Proc. Natl. Acad. Sci. U. S. A. 97, 32363241
7 Duesberg, P. et al. (2000) Explaining high mutation rates of cancer cells
to drug and multidrug resistance by chromosome reassortments that
are catalysed by aneuploidy. Proc. Natl. Acad. Sci. U. S. A. 97,
1429514300
8 Shih, I.M. et al. (2001) Evidence that genetic instability occurs at an
early stage of colorectal tumorigenesis. Cancer Res. 61, 818822
9 Saunders, W.S. et al. (2000) Chromosomal instability and cytoskeletal
defects in oral cancer. Proc. Natl. Acad. Sci. U. S. A. 97, 303308
10 Gisselson, D. et al. (2001) a Abnormal nuclear shape in solid tumours
reflects mitotic instability. Am. J. Pathol. 158, 199206
11 Schmutte, C. and Jones, P.A. (1998) Involvement of DNA methylation
in human carcinogenesis. Biol. Chem. 379, 377388
12 Cremer, T. and Cremer, C. (2001) Chromosome territories, nuclear
architecture and gene regulation in mammalian cells. Nat. Rev. Genet. 2,
292301
13 Bardelli, A. et al. (2001) Carcinogen-specific induction of genetic
instability. Proc. Natl. Acad. Sci. U. S. A. 98, 57705775
14 Jallelapali, P.V. and Lengauer, C. (2001) Chromosome segregation and
cancer: cutting through the mystery. Nat. Rev. Cancer 1, 109117
15 Griffin, C.S. et al. (2000) Mammalian recombination-repair genes
XRCC2 and XRCC3 promote correct chromosome segregation.
Nat. Cell Biol. 2, 757761
1136
www.drugdiscoverytoday.com
research focus
reviews
www.drugdiscoverytoday.com
1137