Mol Biol Rep (2013) 40:66576664

DOI 10.1007/s11033-013-2780-3

Interaction between obesity-related genes, FTO and MC4R,

associated to an increase of breast cancer risk
Patrcia Amorim da Cunha Lia Kubelka de Carlos Back
Aline Fernanda Rodrigues Sereia Clara Kubelka
Maria Cecia Menks Ribeiro Braulio Leal Fernandes
Ilada Rainha de Souza

Received: 19 February 2013 / Accepted: 14 September 2013 / Published online: 4 October 2013
Springer Science+Business Media Dordrecht 2013

Abstract Breast cancer (BC) is a complex disease and

obesity is a well-known risk factor for its development,
especially after menopause. Several studies have shown
Single Nucleotide Polymorphisms (SNPs) linked to overweight and obesity, such as: rs1121980 (T/C) and
rs9939609 (A/T) in Fat Mass and Obesity Associated gene
(FTO) and rs17782313 (T/C) in Melanocortin 4 Receptor
gene (MC4R). Thus, we aimed to investigate the association between these obesity-related SNPs and BC risk. One
hundred BC patients and 148 healthy women from Santa

P. A. da Cunha, L. K. de Carlos Back and A. F. R. Sereia contributed

equally to this work.
P. A. da Cunha
L. K. de Carlos Back

M. C. M. Ribeiro
I. R. de Souza
Cell Biology, Embriology and Genetics Department (UFSC,
BEG), Federal University of Santa Catarina, Florianopolis,
Brazil
P. A. da Cunha
L. K. de Carlos Back

A. F. R. Sereia
C. Kubelka
Biogenetika Diagnostico Molecular e Medicina Genomica,
Florianopolis, Brazil
B. L. Fernandes
Federal University of Santa Catarina, University Hospital (HUUFSC), Florianopolis, Brazil
B. L. Fernandes
Carmela Dutra Maternity Hospital, Florianopolis, Brazil
I. R. de Souza (&)
Laboratorio de Polimorfismos Geneticos, Depto. BEG, Centro de
Ciencias Biologicas, Universidade Federal de Santa Catarina,
Sala 311, Campus Universitario Setor F ed. Fritz Muller,
Trindade, Mailbox: 476, Florianopolis, SC 88040-900, Brazil
e-mail: rainha@ccb.ufsc.br; iliada.rainha@ufsc.br

Catarina, Brazil entered the study. SNPs were genotyped

using Taqman assays. For statistical analyses SNPStats and
SPSS softwares were used. Association analyses were
performed by logistic regression and were adjusted for age
and Body mass index (BMI). Multiple SNPs inheritance
models (log-additive, dominant, recessive, codominant)
were performed to determine odds ratios (ORs), assuming
95 % confidence interval (CI) and P value = 0.05 as the
significance limit. When analyzed alone, FTO rs1121980
and rs9939609 did not show significant associations with
BC development, however MC4R rs17782313 showed
increased risk for BC even after adjustments (Pvalue = 0.032). Interestingly, the interaction of FTO and
MC4R polymorphisms showed a powerful association with
BC. We observed a 4.59-fold increased risk for woman
who have the allele combination C/T/C (FTO rs1121980/
FTO rs9939609/MC4R rs17782313) (P-value = 0.0011,
adjusted for age and BMI). We found important and
unpublished associations between these obesity-related
genes and BC risk. These associations seem to be independent of their effect on BMI, indicating a direct role of
the interaction between FTO and MC4R polymorphisms in
BC development.
Keywords Breast cancer, FTO
MC4R

Obesity-related genes
Gene interaction
SNPs
Abbreviations
FFA
Free fatty acids
TNFa
Tumour-necrosis factor-a
IGF1
Insulin-like growth factor 1
SHBG
Sex-hormone-binding globulin
GWAS
Genome wide association study
SNP
Single nucleotide polymorphism
BMI
Body mass index

123

6658

HU-UFSC
MCD
CONEP
CEP-HU
CEP-MCD
DNA
PCR
HWE
OR
CI
AIC
BIC

Mol Biol Rep (2013) 40:66576664

University Hospital of Federal University of

Santa Catarina
Carmela Dutra Maternity Hospital
National Ethics Committee for Research
University Hospital Ethical Committee for
Research
Carmela Dutra Maternity Hospital Ethical
Committee for Research
Deoxyribonucleic acid
Polymerase chain reaction
HardyWeinberg Equilibrium
Odds ratio
Confidence interval
Akaike Information Criteria
Bayesian Information Criteria

Human genes
FTO
Fat mass and obesity associated
MC4R Melanocortin 4 receptor

Introduction
Breast cancer (BC) is the most common type of cancer in
women worldwide. It is also a leading cause of cancer
death among women in both developed and developing
regions [1]. Breast cancer is a heterogeneous disease with a
complex inheritance pattern. Many risk factors are
involved in the genesis and in the disease progression.
Some risk factors are well-known for BC, such as the
hormone-related factors (age at menarche, number of
pregnancies and age at menopause onset). The studies
suggest that the high circulating serum concentration of
estrogen hormones is associated to BC due to stimulation
of cell proliferation, which increases the chance of mutations and also the growth of early tumors [2].
Obesity is one of the risk factors for BC, especially after
menopause. Among post-menopausal women, obese subjects
have estrogen levels 50100 % higher than normal-weight
women, which is associated with a more than twofold increase
in the risk for BC [35]. This difference in estrogen levels may
be explained by the fact that before menopause, most of the
oestradiol production is performed in the ovary and is regulated by negative feedbak, which prevents an overproduction
of estrogens in pre-menopausal women. In contrast, after
menopause oestradiol is largely produced in the adipose tissue, where the synthesis is not controlled by any feedback
mechanism. Thus, obese post-menopausal women produce
more estrogen hormones than normal-weight women, which
is associated with the pathological process of BC [2, 6, 7].
Besides increased estrogen levels, other mechanisms can
link adiposity to BC risk. In obese patients, the adipose
tissue releases increased levels of free fatty acids (FFA),

123

tumour-necrosis factor-a (TNFa) and resistin, in addition

there is a reduced level of adiponectin, which leads to the
development of insulin resistance and compensatory
chronic hyperinsulinaemia. High levels of insulin increase
levels of bioavailable insulin-like growth factor 1 (IGF1).
The resulting hyperinsulinaemia associated with high levels of IGF1 promotes cellular proliferation and inhibits
apoptosis in target cells from many tissue types, contributing to tumorigenesis [6, 8].
Increased circulating concentrations of insulin and IGF1
cause a reduction in circulating levels of sex-hormone-binding globulin (SHBG), the major carrier protein for oestradiol.
Lower levels of SHBG increase the free form of oestradiol in
blood, which may contribute to the risk of BC [68].
Genetic factors have an important role in complex diseases. Recent genome wide association studies (GWAS)
have identified genetic variants related to obesity and
overweight, such as the polymorphisms in FTO (fat mass
and obesity associated) and MC4R (melanocortin 4 receptor) genes [911]. The Single Nucleotide Polymorphisms
(SNPs) rs17782313 (T[C) located 188 kb downstream of
the MC4R gene, rs9939609 (T[A) and rs1121980 (C[T) in
the first intron of FTO gene, have shown an effect on Body
Mass Index (BMI) in different populations [9, 11, 12].
Given the association between obesity and BC, the aim
of this study is to evaluate the role of these obesity-related
SNPs on increased BC risk. Our hypothesis is that these
SNPs could have a direct role in the development of BC or
could influence its development through their effect in BMI
and adiposity.

Materials and methods

Subjects
This casecontrol study was performed with a group of 248
women from Santa Catarina, Southern Brazil. The case
group is composed of 100 BC patients, diagnosed at the
University Hospital of the Federal University of Santa
Catarina (HU-UFSC) or Carmela Dutra Maternity Hospital
(MCD). The samples were obtained prior to any radio or
chemotherapy regimen. The control group consists of a
total of 148 healthy women, with no first-degree family
history of BC, and was obtained from voluntary groups of
HU-UFSC. All women were informed about the study
objectives, confidentiality of information, and signed a
term of informed consent, allowing the use of their biological material and their epidemiological and clinical data.
The study was approved by national and institutional ethical committees for research (CONEP, CEP-HU and CEPMCD). Epidemiological and clinical data were obtained
based on interviews and medical records.

Mol Biol Rep (2013) 40:66576664

Genotyping
Genomic DNA was extracted from peripheral blood leukocytes by the phenolchloroform method. The genotypes
of the three SNPsMC4R rs17782313 (T[C), FTO
rs1121980 (C[T) and FTO rs9939609 (T[A)were
determined by using StepOne Real-Time PCR System.
Genotyping was carried out using validated TaqMan allelic
discrimination assays (ID C_32667060_10 for rs17782313,
C_2031261_10 for rs1121980 and C_30090620_10 for
rs9939609Applied Biosystems, Foster City, CA, USA).
Genotypes were read by automated software (StepOne
Software version 2.2.2, Applied Biosystems, Foster City,
CA, USA). Reactions were run in 20 lL volumes using an
amplification protocol of 60 C for 30 s, 95 C for 10 min,
followed by 50 cycles of 92 C for 15 s. Lastly a cycle of
60 C for 1 min and 30 s and 60 C for 30 s was performed. Negative controls were included in each plate.
Statistical analysis
All statistical analyses were performed with either SNPStats (available at http://bioinfo.iconcologia.net/index.
php?module=Snpstats) or SPSS (version 12.0; Chicago,
IL). Genotype and allele frequencies were assessed and
tested for HardyWeinberg Equilibrium (HWE). Quantitative and categorical variables were compared using
analysis of Students t test, Persons Chi square or Fisher
Exact Test. The association analyses were performed by
logistic regression and were adjusted for age and BMI.
Multiple SNPs inheritance models (log-additive, dominant,
recessive and codominant) were performed to determine
odds ratios (ORs), assuming 95 % confidence interval (CI)
and P value B0.05 was considered the significance limit.
Using Akaike Information Criteria (AIC) and Bayesian
Information Criteria (BIC) we selected the best SNP
inheritance model. The linkage disequilibrium between the
FTO SNPs was measured using D and r2 statistics. We
estimated the frequencies of allele combinations in the
three different loci and their association with BC. The
statistical power of this association was tested with the
Power Calculator for Casecontrol Genetic Association
Analyses (PGA) [13].
Results
Descriptive statistics is summarized in Table 1. No significant differences were observed between cases and
controls ancestry. Most of the individuals are from European ancestry, which is consistent with the colonization
history of Santa Catarina state. The mean ages did not
differ statistically between cases (53.89) and controls
(52.11) (P value = 0.304). There was no significant

6659
Table 1 Descriptive statistics of ancestry, age, anthropometric
measures and menopausal status in cases and controls
Cases n = 100

n (%)

Controls
n = 148
n (%)

European ancestry

80 (80 %)

121 (82 %)

0.729a

African ancestry

13 (13 %)

17 (11 %)

0.720a

Indigenous ancestry

2 (2 %)

2 (1 %)

0.531b

Asian ancestry

1 (1 %)

0 (0 %)

0.403b

Unknown ancestry

4 (4 %)

8 (5 %)

0.426b

Age (y): mean (range)

53.89 (2486)

52.11 (2580)

0.304c

Weight (kg): mean

(range)

67.79
(40104)

67.14
(45106)

0.677c

Height (cm): mean

(range)

158 (138176)

160 (140180)

0.123c

BMI (kg/m2): mean

(range)

27.16
(17.344.2)

26.37
(1854.1)

0.227c

Normal weight (BMI

\25)

37 (37 %)

64 (43 %)

Obese/overweight (BMI
C25)

63 (63 %)

84 (57 %)

0.326a

Overweight (25B BMI

\30)

41 (41 %)

58 (39 %)

0.775a

Obese (BMI C30)

22 (22 %)

26 (18 %)

0.386a

Pre-menopause

42 (42 %)

40 (27 %)

0.059a

Post-menopause

58 (58 %)

93 (63 %)

Missing

0 (0 %)

15 (10 %)

Pearsons Chi square P value

Fishers exact test P value

T-test P value

difference in weight, height and BMI mean between the

two groups. Subjects were organized according to their
BMI and gathered into three groups: obese (BMI C30),
overweight (25B BMI \30) and obese/overweight (BMI
C25) subjects. The individuals were also classified by
menopausal status. The percentages of subjects in each
group did not differ statistically among cases and controls.
The clinic pathologic features of patients are detailed in
Table 2.
No significant deviations from HWE were found for the
three SNPs, with P values in cases and controls, respectively of: FTO rs11219800.84/0.62, FTO rs9939609
0.67/0.31, MC4R rs177823130.78/1.00. This is one of
the first studies to determine the allele frequencies of these
FTO and MC4R SNPs in a Brazilian population. We found
minor allele frequencies in cases and controls, respectively:
FTO rs1121980 (T)0.40/0.45, FTO rs9939609 (A)
0.38/0.41, MC4R rs17782313 (C)0.23/0.15. Genotype
frequencies and association analyses of the SNPs and BC
are detailed in Table 3. FTO SNPs rs1121980 and
rs9939609 did not show any significant association with

123

3.711 (0.080)

1.583 (0.110)

0.818 (0.634)

n = 100

0.856 (0.588)

Clinicopathologic information

0.850 (0.579)

Table 2 Clinic pathologic features of the 100 patients of breast

cancer

0.649 (0.260)

Mol Biol Rep (2013) 40:66576664

OR2 (P value)

6660

123

3.567 (0.080)

1.596 (0.102)

2.838 (0.154)
3.085 (0.117)

0.856 (0.679)

1.740* (0.044)

1.712 (0.051)

0.836 (0.512)

* Odds ratio with significant P values (B0.05)

60 (60 %)
TT

107 (72 %)

1.700* (0.024)
38 (26 %)
34 (34 %)
TC

48 (32 %)

3 (2 %)

OR1 odds ratio of crude analysis, OR2 odds ratio adjusted for age and BMI

1.664* (0.032)

0.817 (0.457)
37 (37 %)

6 (6 %)

0.767 (0.519)

0.832 (0.516)

0.858 (0.687)

0.622 (0.208)

0.835 (0.534)

OR1 (P value)
OR2 (P value)

0.706 (0.305)
0.690 (0.273)
0.788 (0.386)

0.875 (0.498)
TT

CC
MC4R/rs17782313

BC. However, MC4R rs17782313 SNP presented some

important associations. In the crude analysis of dominant
model we found a significant association between MC4R
rs17782313 and BC in which OR was 1.740, with CI 95 %
of 1.0162.980 and P value of 0.044. Nevertheless this
association was lost when the subjects data were adjusted
for age and BMI. Another significant association between
MC4R rs177823 and increased risk for BC was found in
the log-additive model, which remained even after
adjustments for age and BMI (OR = 1.664, CI 95 %
1.0452.650, P value = 0.032).
By dividing the subjects into pre and post-menopausal
groups we aimed to evaluate whether the effect of these
SNPs in BC risk is influenced according to menopausal

0.864 (0.456)

25

22 (15 %)

53

Unknown

78 (53 %)

22

Negative

50 (50 %)

Positive

AT

23

HER2/neu

46 (31 %)

29

Unknown

13 (13 %)

Negative

37 (37 %)

48

AA

Positive

CC

21

Progesterone receptor (PR)

FTO/rs9939609

20

Unknown

0.768 (0.333)

59

Negative

0.810 (0.253)

22

Estrogen receptor (ER)

Positive

0.795 (0.212)

34

Unknown

70 (47 %)

44

Negative

32 (22 %)

Positive

47 (47 %)

Lymph node involvement

16 (16 %)

7
16

TC

Unknown

TT

33

T3

FTO/rs1121980

T2

OR2 (P value)

37

OR2 (P value)

T1

OR1 (P value)

Tis

Cases
(n = 100)

Tumor size

Gene/SNP ID

30

Dominant

29

Unknown

Log-Additive

Frequency No. (%)

32

Recessive

Genotype

Elston grade
1

Reference

Unknown

1
17

Table 3 Genotype frequencies and association analyses between breast cancer and FTO rs1121980, FTO rs9939609 and MC4R rs17782313 SNPs

Others

OR1 (P value)

IDC and ILC

OR1 (P value)

66

ILCinvasive lobular carcinoma

Controls
(n = 148)

IDCinvasive ductal carcinoma

Codominant

Tumor type

1.540 (0.298)

2.633 (0.218)
3.333 (0.112)
2.069 (0.347)

0.614 (0.363)

2.830 (0.166)

0.651 (0.409)

2.095* (0.044)

1.648 (0.196)

0.550 (0.103)
0.598 (0.139)

* Odds ratio with significant P values (B0.05)

OR1 odds ratio of crude analysis, OR2 odds ratio adjusted for age and BMI

36 (62 %)
TT

72 (77.5 %)

17 (29 %)
TC

29 (31 %)

18 (19.5 %)

5 (9 %)

25 (43 %)

CC
MC4R/rs17782313

TT

3 (3 %)

1.856* (0.035)

1.529 (0.166)

0.639 (0.100)
0.680 (0.136)
50 (54 %)
27 (47 %)
AT

26 (28 %)

14 (15 %)
6 (10 %)

24 (41 %)
CC

AA
FTO/rs9939609

1.889 (0.107)

0.576 (0.150)

0.405 (0.143)
0.497 (0.211)

0.626 (0.197)

0.586 (0.174)

0.303* (0.033)
0.345* (0.040)

0.650 (0.249)

0.450 (0.097)
0.443 (0.084)
0.502 (0.064)
0.550 (0.090)
0.576* (0.032)
0.600* (0.037)
7 (12 %)

27 (47 %)

TT

TC

22 (24 %)
FTO/rs1121980

45 (48 %)

OR1 (P value)
OR1 (P value)
Controls
(n = 93)
Cases
(n = 58)
Gene/SNP ID

OR2 (P value)

OR1 (P value)

OR2 (P value)

OR1 (P value)

OR2 (P value)

Codominant
Recessive
Dominant
Log-Additive
Frequency No. (%)
Genotype

Obesity is a major public health problem, according to the

Brazilian Institute of Geography and Statistics (IBGE),
approximately 40 % of the Brazilian population is overweight. The Household Budget Survey of 20022003
(POF) estimated that 13.1 % of the Brazilian women are
obese. The Brazilian Southern region presents the highest
rates of female obesity (15.1 %) and also one of the highest
rates for BC [14, 15].
Obesity and overweight are important risk factors for
BC development. Many potential mechanisms that can
induce cell proliferation have been proposed to justify this
increased risk, among them we can highlight: high circulating serum concentration of estrogen hormones (mainly
in post-menopausal women), insulin resistance and
hyperinsulinaemia.
Breast cancer is a disorder with a complex inheritance
pattern. The investigation of different genetic variants can
lead to identification of new pathways and consequently
novel preventive and therapeutic targets to improve early

Reference

Discussion

Table 4 Association analyses between breast cancer and FTO rs1121980, FTO rs9939609 and MC4R rs17782313 SNPs in post-menopausal women

status. In pre-menopausal women, FTO rs1121980, FTO

rs9939609 and MC4R rs17782313 showed no association
with BC. In post-menopausal women (Table 4), FTO
rs1121980 showed a protection for BC. This protection
was observed in the log-additive model, even after
adjustments for age and BMI (OR = 0.576, CI 95 %
0.3480.953, P value = 0.032) and in the codominant
model, where the TT genotype presented a protection for
BC also in the adjusted analysis (OR = 0.303, CI 95 %
0.1010.911, P value = 0.033). In addition MC4R
rs17782313 showed increased risk for BC in the logadditive model (OR = 1.856, CI 95 % 1.0463.292,
P value = 0.035) and in the dominant model
(OR = 2.095, CI 95 % 1.0204.302, P value = 0.044),
although these associations were lost after adjustments for
age and BMI.
The log-additive model showed the lowest AIC and BIC
values in all the analyses for the three SNPs. Therefore, it
was selected as the best model to explain the inheritance
pattern of these polymorphisms.
Strong linkage disequilibrium between FTO rs11219890
and FTO rs9939609 SNPs was found: D = 0.9538 and
r2 = 0.8961. The allele combinations frequencies of the
three SNPs were estimated in case and control groups and
their association with BC was calculated (Table 5). The
allele combination C/T/C, for FTO rs1121980, FTO
rs9939609 and MC4R rs17782313 respectively, presented
a strong and powerful association with BC (OR = 4.56, CI
95 % 1.8311.35, P value = 0.0013), with BMI and age
adjustments. The statistical power of this association was
greater than 80 %, the limit for biological studies.

6661
OR2 (P value)

Mol Biol Rep (2013) 40:66576664

123

6662

diagnosis and life expectancy for these patients. The role of

obesity-related genes polymorphisms in BC development is
still uncertain and there are few studies in this field [16,
17]. We selected three obesity susceptibility SNPs, two in
FTO gene (rs1121980 and rs9939609) and one in MC4R
gene (rs17782313) to investigate a possible association
with BC. The study was directed toward investigating
whether these polymorphisms are associated with BC onset
in an indirect or direct manner. The indirect pathway would
be by contributing to obesity and overweight development
and consequently increasing adiposity, estrogen and insulin
levels, which would support tumor growth. The other
hypothesis is that these genetic variants could have a direct
effect in a still unknown pathway related to tumorigenesis
(Fig. 1).
FTO is a large gene with nine exons and eight introns
located in chromosome 16q12.2. The protein encoded by
FTO is a 2-oxoglutarate-dependent enzyme which can be
found in the nucleus. The function of FTO protein is still
uncertain, but it is suggested a role in the maintenance of
genome integrity through repairing or modifying the
nucleic acid [18]. The FTO protein is widely expressed
throughout the body, especially in the hypothalamus nuclei
responsible for energy balance [19, 20]. FTO gene was first
identified in humans in 2007 by Frayling et al. in a GWAS
performed in type 2 diabetes patients. In this study they
identified a common variant in the first intron of FTO (SNP
rs9939609) which was linked to increased BMI measures
of the subjects. Sixteen percent of adults who were
homozygous for the risk allele, weighed about 3 kg more
and had 1.67-fold increased odds of developing obesity
when compared with those not inheriting the risk allele [9].
This association of FTO rs9939609 with increased BMI
was replicated by several studies in different populations
[9, 2128]. There are other SNPs in FTO associated with
elevated BMI. The rs11219890 SNP, also located in the
first intron of FTO, was identified in 2007 by Dina et al.
[12] in a study with French subjects, where they found an
association with severe obesity. Subsequent studies also
found association of FTO rs1121980 with obesity [25, 29
35].
Many studies have demonstrated that FTO rs1121980
and rs9939609 SNPs are in strong linkage disequilibrium,
probably forming haplotypes [24, 25, 29, 33]. Our study
confirmed the linkage disequilibrium between these two
loci in a Brazilian population.
Kaklamani et al. [16] in 2011 reported for the first time
the association between FTO SNPs and BC risk in a NorthAmerican women population. They found that FTO SNPs
are powerful classifiers in predicting BC risk. But in our
study, this association was not replicated. When analyzed
alone, we did not find FTO SNPs being associated with the
risk of BC. When post-menopausal women were analyzed

123

Mol Biol Rep (2013) 40:66576664

separately, FTO rs1121980 showed a protection for BC that

appeared to be independent of age and BMI. This protection
of FTO rs1121980 in post-menopausal women must be
further investigated in other studies, as well as the influence
of this SNP in the proteins function. This SNP could be in a
linkage disequilibrium block with protective genes and that
could justify the protection found in this study.
MC4R gene is located on chromosome 18q22 and it is
composed by a single exon with 1,438 base pairs. This
gene encodes a seven domain-transmembrane G protein
coupled receptor that is expressed in muscle, adipose tissue
and predominantly in the brain [36, 37]. This receptor is
highly expressed in the hypothalamic nuclei, which has
been strongly implicated in the central control of food
intake and energy balance [38]. Mutations in this gene are
well-established as the most frequent cause of monogenic
form of extreme, early-onset obesity [36]. In the last years,
common genetic variation in and near MC4R have been
reported to contribute to common obesity susceptibility
[11]. The SNP rs17782313, located 188 kb downstream of
MC4R was described for the first time in 2008, in a study
with 16,876 European subjects, where it was associated
with increased BMI in adults and children [11]. This
association has been replicated in many studies in different
populations [24, 31, 3942].
To our knowledge, the present study is the first to report
an association between MC4R gene polymorphisms and BC
risk. We found the C allele of MC4R rs17782313 implicated
in BC development independently of age and BMI
(OR = 1.664, CI 95 % 1.0452.650, P value = 0.032).
This association indicates a possible direct role of this
MC4R polymorphism in the development of BC. This role
seems to be independent of its effect on BMI. The association of MC4R rs17782313 and BC needs to be investigated
and elucidated in further studies. When MC4R rs17782313
was analyzed in post-menopausal women, it showed an
increased risk for BC development. However this association was lost after adjustments for age and BMI, which
indicates that in post-menopausal women the association of
MC4R rs17782313 with BC could be mediated through
obesity or overweight effects.
Another study investigated the association between
MC4R rs17782313 and BC risk. Kusinska et al. [17] in
2011 analyzed FTO, MC4R and NRXN3 genes in Polish
women. They found that only NRXN3 gene was associated
with BC, FTO and MC4R genes did not show any significant association with the disease.
As observed in all our association analysis, the logadditive model showed the lowest AIC and BIC values.
This model was considered the best one to explain the
inheritance pattern of FTO and MC4R SNPs assessed in
this study and it should be taken into account for future
analysis.

Mol Biol Rep (2013) 40:66576664

Table 5 Estimated allele
combination frequencies of
FTO rs1121980, FTO
rs9939609 and MC4R rs177823
SNPs and their association with
breast cancer

6663

FTO
rs1121980

FTO
rs9939609

MC4R
rs17782313

Estimated
frequencies

OR

Cases

(95 % CI)

Controls

P value*

0.4216

0.4864

1.00

0.3128

0.3115

1.28 (0.792.07)

0.32

0.1731

0.0502

4.59 (1.8611.32)

0.0011

0.0569

0.0900

0.65 (0.261.63)

0.36

Obesity risk alleles are in bold

0.0253

0.0428

0.69 (0.232.04)

0.5

ND not determined

0.0103

0.0107

1.44 (0.229.29)

0.7

* P value adjusted for age and

BMI

0.0084

ND

ND

Fig. 1 Possible pathways of

association of FTO and MC4R
polymorphisms and breast
cancer: 1 FTO and MC4R
polymorphisms are risk factors
for breast cancer through an
effect in obesity and
overweight. 2 FTO and MC4R
polymorphisms have a direct
association with breast cancer,
independently of their effect on
obesity and overweight

Interestingly the allele combination C/T/C of FTO and

MC4R polymorphisms (rs1121980/rs9939609/rs17782313)
showed a powerful association with BC risk in the present
study. This allele combination is formed by two non-risk
alleles for obesity (FTO rs1121980 and rs9939609) and by
one obesity risk allele (MC4R rs17782313). The estimated
frequencies of the allele combination C/T/C was significantly higher in BC patients than in healthy women. Thus,
according to our study woman who have this allele combination are 4.59-fold more likely to develop BC when
compared to the general population (P value = 0.0011).
This analysis showed statistical power greater than 80 %
with this sample size and was adjusted for age and BMI.
This result indicates that the interaction between the
obesity non-risk alleles of FTO and the obesity risk allele
of MC4R have a direct influence in predisposing to BC.
However, the risk allele combination for obesity (T/A/C)
did not show increased risk for BC, it was rather protective,
but this analysis was not significant statistically. This is the
first study to report the association of this allele combination (C/T/C) with BC risk. We suggest more studies in
different populations to elucidate the causes and mechanisms involved in this pathway to improve knowledge
about the subject.
In conclusion, our study showed important and unpublished association of FTO and MC4R polymorphisms

interaction with BC. More studies are important to identify

novel disease susceptibility loci for BC, which might
contribute to the development of new prevention strategies,
early detection and more specific treatment approaches.
Acknowledgments The authors are grateful to the volunteers, the
hospitals and the research teams. We also wish to thank the Brazilian
National Council for Scientific and Technological Development
(CNPq), Coordenacao de Aperfeicoamento de Pessoal de Nvel
Superior (CAPES), Fundacao de Amparo a` Pesquisa e Inovacao do
Estado de Santa Catarina (FAPESC) and the Federal University of
Santa Catarina.

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