Temporal Variation and Identification of Factors
Associated with Endogenous Retinoic Acid
Isomers in Serum from Brazilian Women
Erin M. Siegel,
1
Neal E. Craft,
2
Denise J. Roe,
1
Eliane Duarte-Franco,
4
Luisa L. Villa,
3
Eduardo L. Franco,
4,5
and Anna R. Giuliano
1
1
Cancer Prevention and Control Program, University of Arizona Cancer Center and Epidemiology and Biostatistics Division, Mel and Enid
Zuckerman Arizona College of Public Health, Tucson, Arizona;
2
Craft Technologies, Inc., Wilson, North Carolina;
3
Ludwig Institute
for Cancer Research, Sa ˜o Paulo, Brazil; and Departments of
4
Oncology and
5
Epidemiology and Biostatistics, McGill University,
Montreal, Quebec, Canada
Abstract
Objective: Retinoids (natural and synthetic derivatives
of vitamin A) have cancer chemotherapeutic and
chemopreventive activities. Retinoic acid (RA) treat-
ment has been associated with significant regression of
preneoplastic lesions. However, serious toxicity asso-
ciated with some therapies has made long-term che-
moprevention in healthy populations unfeasible.
Recently, serum RA has been shown to increase in re-
sponse to oral retinol (vitamin A) supplementation.
Here, we assess the variability of circulating RA levels
and the lifestyle, demographic, and nutritional factors
that explain such variability.
Method: Total RA concentration and the concentrations
of RA isomers (all-trans -RA, 13-cis -RA, and 9-cis -RA)
were measured by high-pressure liquid chromatogra-
phy in serum samples obtained 4 months apart from
502 women participating in the Ludwig-McGill Cohort
(Sa ˜o Paulo, Brazil).
Results: The relative abundance of the three RA isomers
was similar for each visit (baseline and month 4), with
13-cis -RA having the highest concentrations followed
by 9-cis -RA and all-trans -RA. The within-person
variability of total RA and individual isomers was
low. Using multivariate logistic regression models
(upper tertile versus low/middle tertile of serum RA),
we found that age, race, oral contraceptive use, total
number of pregnancies, and season of initial blood
draw were significantly associated with at least one
endogenous RA isomer level. All endogenous RA
isomers were positively associated with serum retinol,
B-carotene, and B-cryptoxanthin levels.
Conclusion: These results have implications for the
design of future epidemiologic studies focused on as-
sessing RA-disease association and intervention stud-
ies aimed at modulating RA levels. (Cancer Epidemiol
Biomarkers Prev 2004;13(11):1693 – 703)
Introduction
Retinoic acid (RA) is required for many biological
processes including vision, development, and reproduc-
tion (1-3). RA mediates these activities by binding to
nuclear retinoid receptors, members of the steroid hor-
mone receptor superfamily (4), and altering transcrip-
tional activity (1, 5). To date, two families of receptors
have been identified, RA receptor (RAR) and retinoid X
receptor (5). RAR binds all-trans -RA and 9-cis -RA with
equal affinity, whereas retinoid X receptor preferentially
binds 9-cis -RA (6). Each receptor family has at least three
subtypes (a, h, and g), and within these subtypes, there
are many different isoforms (7). Due to the different
receptor isoforms, retinoid receptors can interact with a
diverse number of receptors forming homodimers and
heterodimers not only with retinoid receptors but also
with other steroid hormone nuclear receptors such as
vitamin D receptors and the estrogen receptor (8).
Through this potent activity as transcriptional regu-
lators, retinoids (natural and synthetic derivatives of
vitamin A) have been shown to be cancer chemothera-
peutic and chemopreventive (see also refs. 5, 9, 10 for
reviews). RA treatment (all-trans -RA) has been associated
with significant regression of several preneoplastic
lesions, including oral leukoplakia (11), cervical dysplasia
(12), and actinic keratoses (5). Although retinoid chemo-
prevention is effective, there are toxicities associated with
therapy such as varying degrees of teratogenicity and
mucocutaneous cytotoxicity (5, 13). These toxicities make
long-term chemoprevention among healthy populations
with naturally occurring RA unfeasible. 13-Cis -RA (iso-
tretinoin) and N-(4-hydroxyphenyl)retinamide (Fenreti-
nide) have been shown to have lower toxicities, but it
remains unclear if these drugs are able to induce retinoid
receptor – mediated changes. An alternative retinoid che-
moprevention approach is to develop a method for
increasing endogenous concentrations of RA. Such an
Received 3/5/04; revised 5/6/04; accepted 6/3/04.
Grant support: National Cancer Institute grants CA70269 and CA81310, Canadian
Institutes of Health Research grants MA-13647 and MOP-49396, Ludwig Institute
for Cancer Research intramural grant, Canadian Institutes of Health Research
Distinguished Scientist Award (E.L. Franco), and National Cancer Institute Cancer
Prevention and Control predoctoral fellowship grant R25CA078447 (E.M. Siegel).
The costs of publication of this article were defrayed in part by the payment of
page charges. This article must therefore be hereby marked advertisement in
accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Note: Presented in part at the AACR Annual Conference, San Francisco, California,
2002 and AACR Frontiers in Cancer Prevention Conference, Phoenix, Arizona, 2003.
Requests for reprints: Anna R. Giuliano, H. Lee, Moffitt Cancer Center and Research
Institute, 12902 Magnolia Drive, MRC-2E, Tampa, FL 33612. Phone: 813-903-6820.
E-mail: guiliano@moffitt.usf.edu
Copyright D 2004 American Association for Cancer Research.
Cancer Epidemiology, Biomarkers & Prevention 1693
Cancer Epidemiol Biomarkers Prev 2004;13(11). November 2004
Research.
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