Blood Cells, Molecules, and Diseases 33 (2004) 326 – 329

www.elsevier.com/locate/ybcmd

Cytochrome P4501A1 polymorphism is associated with susceptibility to

acute lymphoblastic leukemia in adult Mexican patients
M.P. Gallegos-Arreolaa,*, C.M. Batista-Gonzáleza,b, J.L. Delgado-Lamasc, L.E. Figuerad,
A.M. Puebla-Péreza, L. Arnaud-Lópeza,e, V. Peralta-Leala,e,
L.J. Ramı́rez-Jiranoa,e, G.M. Zúñiga-Gonzáleza
a
División de Medicina Molecular, Centro de Investigación Biomédica de Occidente, Guadalajara, Jalisco, México
b
Doctorado en Biologı́a Molecular, Centro Universitario de Ciencias de la Salud, Universidad de Guadalajara, Jalisco, México
c
Servicio de Hematologı́a, Hospital de Especialidades, Centro Médico Nacional de Occidente, Guadalajara, Jalisco, México
d
División de Genética, Centro de Investigación Biomédica de Occidente, Guadalajara, Jalisco, México
e
Doctorado en Genética Humana, Centro Universitario de Ciencias de la Salud, Universidad de Guadalajara, Jalisco, México
Submitted 19 May 2004; revised 2 July 2004
Available online 8 September 2004
(Communicated by E. Beutler, M.D., 2 July 2004)

Abstract

We studied the role of cytochrome P4501A1 (CYP1A1 Val/Val) genotypes in the etiology of acute lymphoblastic leukemia (ALL) in adult
Mexican patients. Distributions of CYP1A1 Val/Val genotypes in peripheral blood DNA samples from 136 healthy controls and 136 adult
patients with ALL were evaluated. There was an increased frequency of the CYP1A1 Val/Val genotype among ALL patients, showing a
significant association between this genotype and the risk of developing ALL.
D 2004 Elsevier Inc. All rights reserved.

Keywords: Polymorphism; CYP1A1; Acute lymphoblastic leukemia; Mexican population

Introduction although the clinical, pathological, and immunophenotypic

The etiology of most types of leukemia remains
unknown. Acute lymphoblastic leukemia (ALL) is a
heterogeneous disease characterized by the predominance
of lymphoblasts or immature hematopoietic precursors, in
which the malignant cells express diverse phenotypes with
variable response to chemotherapy.
Despite much research, little is known about incitants of
leukemogenesis, particularly with respect to issues such as
the role of genetic susceptibility and environmental factors,
* Corresponding author. División de Medicina Molecular, Centro de
Investigación Biomédica de Occidente, IMSS Sierra Mojada No. 800,
Colonia Independencia, C.P 44340, Guadalajara, Jalisco, México. Fax: +52
33 3618 17 56.
E-mail address: marthaga@foreigner.class.udg.mx
(M.P. Gallegos-Arreola).
1079-9796/$ - see front matter D 2004 Elsevier Inc. All rights reserved.
doi:10.1016/j.bcmd.2004.07.002
features of the disease are well documented [1,2]. To date,
epidemiological studies have failed to find any reproducible
significant association between ALL and either genetic or
environmental factors [2–4]. It has been suggested that
individuals possessing a modified ability to metabolize
carcinogens are at increased risk of cancer [5,6]. Thus,
polymorphisms in genes encoding carcinogen-metabolizing
enzymes may be relevant in determining susceptibility to
cancer, in that individuals carrying the more active form of
an enzyme involved in the activation of carcinogens or less
efficient alleles of detoxifying enzymes might be at greater
risk of cancer [7,8].
For this reason, the family of genes for phase I
cytochrome P450 has attracted interest. Genetic variants
have been described in Caucasians for the CYP1A1 gene.
Several studies have reported associations between this
variant and altered risk of a variety of cancers, including
 M.P. Gallegos-Arreola et al. / Blood Cells, Molecules, and Diseases 33 (2004) 326–329 327

Table 1 passive smokers, and ex-smokers. The amount of tobacco

Characteristics of the study groups smoke exposure was calculated as pack-years [packs (20
Controls ALL patients cigarettes) per day per year of smoking].
(n = 136) (n = 136)
Genomic DNA was extracted from blood samples
Age (years) according to standard protocols [17]. Histological data were
Mean (SD) 41.9 (2.9) 37.5 (16.1)
obtained from the Service of Hematology, Centro Médico
Range 18–72 18–78
Nacional de Occidente, Instituto Mexicano del Seguro
Gender, n (%) Social.
Female 70 (51%) 79 (58%) Presence of the Val allele in exon 7 of the CYP1A1 gene
Male 66 (49%) 57 (42%) was determined by DNA amplification in a total PCR volume
of 25 AL containing 400 AM dNTPs, 20 pmol of primers, 1.5
Smoking status, n (%)
Ever smoked 63 (46%) 57 (42%) mM MgCl2, and 2.5 U Taq polymerase (Invitrogen, life
Never smoked technologies). We used the following primers: 5V-CCA GGA
Nonpassive smoking 24 (18%) 36 (26%) AGA GAA AGA CCT CCC AGC GGG CCA-3V and 5V-
Passive smoking 49 (36%) 43 (32%) GAA AGG CTG GGT CCA CCC TCT-3V. These primers
amplify part of intron 6 and exon 7 of the human cDNA
CYP1A1 gene, yielding a 263-bp fragment. The PCR
those of the lung, bladder, gastrointestinal tract, skin, cervix, conditions consisted of an initial melting temperature of
and breast [9–15]. The prevalence of this polymorphism 948C (4 min) followed by 35 cycles of melting (948C, 1 min),
varies greatly among different ethnic groups, as well as annealing (688C, 1 min), and extension (728C, 4 min). The
within the Caucasian population [16]. Population data amplified product was subjected to restriction enzyme
should be regarded cautiously because intergroup variation analysis with NcoI (New England Biolabs, Beverly, MA,
can influence the power and interpretation of epidemiolog- USA), according to the manufacturer’s instructions. The
ical data, particularly in a very heterogeneous population samples were separated using 6% polyacrylamide gel
such as ours in Mexico. electrophoresis (29:1) followed by silver staining [18,19].
We used a genotyping approach based on the polymerase This revealed the 232- and 31-bp fragments for wild-type
chain reaction (PCR) to examine genetic polymorphism in alleles (isoleucine) or a single 263-bp fragment when the
CYP1A1 Val/Val in 136 patients with ALL and in 136 mutation (valine) was present.
controls. The frequency of the val/val genotype among ALL
patients compared with controls was tested for significance
by Chi-squared tests with Yates correction. Odds ratios and
Materials and methods 95% confidence intervals (CI) were also calculated. Two-
sided P b 0.05 was assumed to be statistically significant.
We recruited 136 healthy donors and 136 patients with
ALL from Mexicans living in Guadalajara City and its
surroundings. Patients were recruited from June to Decem- Results and discussion
ber 2003. Controls were recruited from volunteer blood
donors, and their habits and family histories were inves- Among the ALL patients, the average age was 37.5 years
tigated. All individuals signed a letter of informed consent. and 58% were female. In the control group, the average age
Both groups were interviewed for personal data collec- was 41.9 years and 51% were female (Table 1).
tion with a short questionnaire that included items on age, Table 2 shows the number of cases and controls by
gender, detailed information on recent and past tobacco use, CYP1A1 status. The frequency of CYP1A1 Val allele was 6%
alcohol and any other type of drug consumption, occupa- (8 of 136) among the population controls and 16% (22 of 136)
tional exposure, and any history of the use of chemotherapy among the ALL patients. The ALL patients showed a
agents in the case of the ALL patients. The subjects were significantly higher frequency of Val/Val genotype, with an
classified as smokers (z3 cigarettes per day), nonsmokers, odds ratio (OR) of 3.0 (95% CI, 1.3–7.2), and Yates-corrected

Table 2
Genotype distribution of CYP1A1 polymorphisms in healthy controls and patients with ALL
Genotype Alleles Controls versus ALL patients
ISO/ISO ISO/Val Val/Val ISO Val OR 95% CI P
n (%) n (%) n (%) 2n (%) 2n (%) Low–high
Controls 69 (51) 59 (43) 8 (6) 197 (72) 75 (28) 3.1 1.3–7.2 0.012
ALL patients 49 (36) 65 (48) 22 (16) 163 (60) 109 (40) 1.8 1.2–2.5 0.003
328 M.P. Gallegos-Arreola et al. / Blood Cells, Molecules, and Diseases 33 (2004) 326–329
Table 3
CYP1A1 a2B genotype frequencies in patients with ALL and healthy
controls
CYP1A1 b Controls, n (%) Cases, n (%) OR (95% CI)
ISO/ISO 69 (48) 49 (36)
ISO/Val 59 (43) 65 (48) 1.0a
Val/Val 8 (9) 22 (16) 3.0 (1.3–7.2)
CI: confidence interval.
a
This reference category combined the two genotypes because the odds
ratio (OR) showed an equal risk for each.
b
The genotype frequency showed Hardy-Weinberg equilibrium.
Chi-squared test showing P b 0.05. The genotype distribu-
tions from cases and controls were all consistent with Hardy-
Weinberg equilibrium according to the likelihood ratio.
Table 3 shows the overall distribution of ALL patients
and controls according to the Val/Val genotype. The odds
ratio was 3.0 (95% CI, 1.3–7.2).
Little attention has been paid to the role of genetic
susceptibility and environmental exposure in the etiology of
ALL. In particular, people with an altered ability to activate
procarcinogens and detoxify carcinogens may have an
increased risk of developing cancer [6]. Indeed, epidemio-
logical studies have shown that the risk of leukemia is
associated with drug use and occupational exposure to
pesticides [20,21]. Our observations support possible genetic
and environmental interactions for the risk of developing
ALL in adults.
We found that 16% of the ALL patients and 9% of
healthy controls had a Val/Val genotype. For cancer screen-
ing, it is important that the fraction of controls with a marker
should be very small to avoid large numbers of unnecessary
interventions [22]. Thus, the CYP1A1 Val/Val phenotype
offers a possibility for early detection of individuals at a
high risk for developing ALL in the Mexican population, if
combined with another marker to improve performance.
Adults carrying the CYP1A1 Val allele thus show a
greater risk of developing ALL (OR, 3.0). To our
knowledge, this is the first report of an association
between a CYP1A1 variant and the risk of developing
ALL in the Mexican population. The CYP1A1 Val allele is
associated with elevated enzymatic activity supporting the
hypothesis of linking the risk of ALL with the inducibility
of the xenobiotic metabolizing enzyme CYP1A1 [23–25].
Consequently, carriers of variant alleles are expected to
present greater risk when exposed to carcinogens such as
polycyclic aromatic hydrocarbons (PAHs) [25]. Indeed,
there is an association between the induction of placental
CYP1A1 activity and PAH in cigarette smoke and cooked
foods [26]. However, the possibility that these results are
due to chance in the context of multiple testing cannot be
ruled out [27,28]. Furthermore, the influence of this
genotype may be more obvious when other variants
participate in the effect. Hormonal factors that play an
obvious role in breast cancer and other sources of variation
in cancer risk due to gender have received little attention
[27,28]. Different studies performed in adults with non-
hematological neoplasms provide further evidence for an
increase in the risk of cancer in patients carrying both
P
CYP1A1- and GSTM1-associated risk alleles [29,30]. This
is supported biologically by a recent study showing that
b0.05
the combination of CYP1A1 and GSTM1 genotypes clearly
affects the formation of DNA adducts in human white
blood cells [31]. Our results are of course limited in
number and require confirmation from larger studies.
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