Original article 1

Melanoma risk is associated with vitamin D receptor gene

polymorphisms
Katarina Zeljica,f,*, Lidija Kandolf-Sekulovicb,e,*, Gordana Supica,e,
Janko Pejovicc, Marijan Novakovicd,e, Zeljko Mijuskovicb,e and Zvonko Magica,e
Previous studies have reported that vitamin D receptor
(VDR) gene polymorphisms are associated with the
occurrence of various cancers, including melanoma. The
aim of the current study was to investigate the association
of VDR gene polymorphisms with melanoma risk,
clinicopathological characteristics, and vitamin D levels.
The study group included 117 patients (84 patients with
superficial spreading melanoma and 33 patients with
nodular melanoma). The control group included 122
sex-matched and age-matched healthy-blood donors of
the same ethnicity. VDR gene polymorphisms FokI, EcoRV,
TaqI, and ApaI were genotyped by real-time PCR. In 60
patients, the total 25-hydroxyvitamin D levels were
evaluated in serum samples by direct chemiluminescence.
Associations among parameters were considered to be
significant if the P value was less than 0.05. Significant
differences in the frequencies of VDR genotypes were
observed between cases and the control group for FokI
and TaqI polymorphisms (P < 0.0001; P = 0.005,
respectively). Heterozygous Ff as well as mutant FF
genotypes of the FokI polymorphism were associated with
increased melanoma risk compared with the wild-type
form [odds ratio (OR) = 3.035, P = 0.003; OR = 9.276,
P < 0.0001, respectively]. A significantly increased
melanoma risk was observed for the heterozygous Tt
(OR = 2.302, P = 0.011) and the mutated variant tt

(OR = 3.697, P = 0.003) of the TaqI polymorphism in

comparison with the wild-type genotype. None of
the polymorphisms studied was associated with
clinicopathological characteristics and vitamin D serum
level. Our results suggest that FokI and TaqI
polymorphisms in the VDR gene may be considered as
potential biomarkers for melanoma susceptibility. Low
vitamin D levels in melanoma patients indicate the need
for vitamin D supplementation. Melanoma Res 00:000000

c 2014 Wolters Kluwer Health | Lippincott Williams &
Wilkins.

Introduction

Numerous studies have shown that vitamin D exerts

antiproliferative, prodifferentiation, proapoptotic, and
angiogenesis inhibitor effects [8,9]. There are also some
data, which require further confirmation, on the use of
vitamin D as a preventive and therapeutic cancer agent,
supported by preclinical and clinical studies in different
cancer types, including melanoma [10].

Cutaneous melanoma, in its advanced stage, is one of the

most aggressive and, despite recent advances in treatment, still incurable diseases, with a median survival of
less than 1 year [13]. The incidence of melanoma is
increasing worldwide [4] and the main risk factors for its
development are ultraviolet radiation, number of nevi,
and skin phototype [5]. Genetic predisposition plays a
significant role in the development of this cancer [57].
Apart from high-penetrance genes such as CDK4 and
CDK6 and low-penetrance genes linked to the phenotype
such as MC1R, well known for their role in melanoma
development, considerable efforts have been made to
discover other potential biomarkers that can be used
reliably to help identify individuals at risk or predict the
disease course.

All supplementary digital content is available directly from the corresponding

author.
c 2014 Wolters Kluwer Health | Lippincott Williams & Wilkins
0960-8931 

Melanoma Research 2014, 00:000000

Keywords: melanoma risk, single nucleotide polymorphisms, VDR gene,
vitamin D level
a

Laboratory for Molecular Genetics, Institute for Medical Research, bClinic for
Dermatovenereology, cCenter for Biochemistry, dClinic for Plastic Surgery and
Burns, Military Medical Academy, eFaculty of Medicine, Military Medical Academy,
University of Defense and fDepartment of Genetics and Evolution, Faculty of
Biology, University of Belgrade, Belgrade, Serbia
Correspondence to Katarina Zeljic, PhD, Laboratory for Molecular Genetics,
Institute for Medical Research, Military Medical Academy, Crnotravska 17,
11 000 Belgrade, Serbia
Tel: + 381 63 84 52 350; fax: + 381 11 266 27 22;
e-mails: katjaze@yahoo.com, katarina.zeljic@bio.bg.ac.rs
*Katarina Zeljic and Lidija Kandolf-Sekulovic contributed equally to the writing of
this article.
Received 30 December 2013 Accepted 11 February 2014

Previous studies have reported the proapoptotic effects

of vitamin D and cell migration inhibition in human
melanoma cell lines in vitro [11]. In melanoma patients
living in areas with low insolation, suboptimal concentrations of vitamin D were linked to greater Breslow
thickness and melanoma progression [12]. Similar results
were also found in areas with high insolation, such as
Spain, where in the majority of melanoma patients at
diagnosis, suboptimal vitamin D concentrations were also
detected [13]. In contrast, in a Danish study, higher
concentrations of vitamin D were found to be a biomarker
for increased sun exposure and risk factor for melanoma
DOI: 10.1097/CMR.0000000000000065

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2 Melanoma Research 2014, Vol 00 No 00

development [14]. No data have been reported on the

vitamin D levels in melanoma patients from southeast
Europe, an area with continental climate, low insolation
during the winter, and high insolation in the summer. In this
study, we report for the first time in southeast Europe
vitamin D levels from our group of melanoma patients.
The vitamin D functions are mediated through the nuclear
vitamin D receptor (VDR), which acts as a ligand-inducible
transcription factor [15]. The VDR protein is coded by the
VDR gene (chromosome location 12q1214) [15], which is
expressed in both normal and malignant melanocytes [3].
Numerous single nucleotide polymorphisms (SNPs) have
been identified in the VDR gene, which might compromise
receptor functionality and consequently contribute toward
cancer. The EcoRV (rs4516035) polymorphism is located in
the VDR gene promoter region without known functional
effects. FokI (rs2228570) is the most studied VDR gene
polymorphism with functional effects. Namely, the mutated
form results in the synthesis of a VDR protein shorter by
three amino acids, which has increased transactivation
capacity in comparison with the wild type [15,16]. ApaI
(rs7975232) and TaqI (rs731236) polymorphisms, located in
intron 8, are silent SNPs found in linkage disequilibrium
(LD) [15]. A number of studies have reported an association
between VDR gene polymorphisms and the risk of
occurrence of various cancers, such as breast [17], colon [18],
and oral cancer [19], but also melanoma [3,13,20]. Despite
the important role of vitamin D in carcinogenesis, genetic
variants in the VDR gene in melanoma have been
investigated in a limited number of studies. As the data
from previous studies are inconsistent, the aim of the current
study was to investigate the association between VDR gene
polymorphisms and the risk of occurrence of melanoma as
well as the association with clinicopathological characteristics,
prognosis, and vitamin D serum levels in melanoma patients
from Serbia.

Patients and methods

Study group and samples

The study group comprised 117 patients (56 men, 61

women, median age 54 years) with newly diagnosed
melanoma in 2011 at the Melanoma Center, Military
Medical Academy, Belgrade, Serbia. Eighty-four patients
had superficial spreading melanoma and 33 patients had
nodular melanoma. Patients with other melanoma subtypes were excluded from the study. Melanoma risk
factors were recorded on the basis of a structured
questionnaire containing data on sun exposure, physician
recorded skin phototype, eye color, number of nevi,
presence and number of solar lentigo, family history of
melanoma and other cancers, and personal history of skin
cancer. Clinicopathological characteristics were retrieved
from the hospital database, and the following parameters
were recorded: age, sex, melanoma subtype, tumor
location, Breslow thickness, ulceration, pT stage, and
clinical stage at diagnosis. Disease staging was performed

according to the current AJCC classification from 2009.

The patients were followed up for a median of 26 months
(343 months).
The control group included 122 healthy-blood donors,
White individuals of the same ethnicity with no personal
cancer history, matched in sex and age. Peripheral blood
samples were collected from both the patient and the
control groups and stored at 201C until DNA isolation.
This study was approved by the Ethics Committee of the
Military Medical Academy, according to the Helsinki
Declaration (2008). All individuals provided informed
consent for the use of biological samples for research
purposes.
DNA isolation

DNA was isolated from the peripheral blood samples

using a Blood Prep kit at the AbiPrism 6100 NucPrep
Station (Applied Biosystems, Foster City, California,
USA) following the manufacturers instructions. DNA
samples were stored at 201C until further analysis.
Genotyping

Polymorphisms EcoRV (rs4516035), FokI (rs2228570),

ApaI (rs7975232), and TaqI (rs731236) in the VDR gene
were genotyped by the allelic discrimination method
using the TaqMan SNP Genotyping Assays (Applied
Biosystems). Data analyses were carried out using SDS
Software on a real-time PCR 7500 system (Applied
Biosystems).
Measurement of serum 25-hydroxyvitamin D

In 60 melanoma patients, the total 25-hydroxyvitamin D

level was measured by direct chemiluminescence (Advia
Centaur XP; Siemens, Erlangen, Germany) in serum
samples collected at diagnosis and up to 6 months after
melanoma diagnosis during the winter months. Insufficient levels of 25-hydroxyvitamin D were defined as less
than 20 ng/ml (< 50 nmol/l) [13].
Statistical analysis

Collected data were analyzed using SPSS software (version

20.00; IBM Inc., Chicago, Illinois, USA). Binary logistic
regression was used for odds ratio (OR) calculation with 95%
confidence interval (CI). All reported ORs and 95% CIs
were adjusted for sex and age. Dominant, recessive, and
log-additive genetic inheritance models were tested. Hazard
ratio was calculated by the Cox proportional hazard regression
analysis with 95% CI. KaplanMeier survival curves were
compared using the log-rank test.
Using Haploview software (version 4.2) [21], pairwise LD
(D0 and r2 values), haplotype frequencies, and departure
from HardyWeinberg equilibrium were determined.
Haplotype block was identified using the option solid
spine block definition. Permutation testing to calculate corrected P values for multiple testing of 1000

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VDR gene polymorphisms in melanoma Zeljic et al.

simulations was performed using Haploview software.

Haplotype association analysis was carried out using
Thesias software (version 3.1) [22]. Calculated P values
were two-tailed and considered significant when P values
were less than 0.05.

Results
Association between vitamin D receptor single
nucleotide polymorphisms, melanoma risk, and
clinicopathological characteristics

Genotype and allele frequencies of polymorphisms

analyzed in the VDR gene are presented in Table 1.
Significant differences in the VDR allele and genotype
frequencies between the case and the control groups
were observed for the FokI and TaqI polymorphisms
(P < 0.0001; P < 0.0001, respectively) (Table 1).
Adjusted OR analysis showed that the heterozygous Ff as
well as the mutated FF genotype of the FokI polymorphism
were associated with an increased risk of melanoma
compared with the wild-type genotype (OR = 2.860,
P = 0.004; OR = 9.033, P < 0.0001, respectively) (Table 2).
Significantly increased risk of melanoma was observed for the
heterozygous Tt (OR = 2.323, P = 0.011) and the mutated
variant tt (OR = 3.558, P = 0.003) of the TaqI polymorphism
compared with the wild-type genotype (Table 2).
Mutated allele carriers (combined heterozygous and mutated
genotypes) were associated with a significantly increased risk
of melanoma compared with the wild-type genotype for the
FokI and TaqI polymorphisms (OR = 4.015, P < 0.0001;
OR = 2.616, P = 0.002, respectively) (Table 2).
None of these polymorphisms was associated with
clinicopathological characteristics: melanoma type, Breslow thickness, ulceration, tumor size, presence of regional
or distant metastases, and stage at diagnosis (Supplementary Table 1). Cox regression analysis showed that
Breslow thickness, tumor size, nodal status, and stage
were significant indicators for overall survival (Table 3).
However, the VDR gene SNPs studied here were not
associated with overall survival (KaplanMeier survival
curves not shown).
Table 1

Haplotype analysis and linkage disequilibrium

Pairwise LD coefficients (D0 and r2 values) for VDR gene

SNPs, on the basis of genotypes of 239 individuals in the
casecontrol study and in each of the case and control
groups separately, were calculated and plotted (Fig. 1).
VDR ApaI and TaqI polymorphisms were in LD in the
cases (D0 = 0.59, r2 = 0.17) and the control group (D0 = 1,
r2 = 0.32), as well as the case and control groups
combined (D0 = 0.78, r2 = 0.24) (Fig. 1).

Association between haplotypes and melanoma risk

and clinicopathological characteristics

In further analyses, we considered haplotypes of VDR

ApaI and TaqI polymorphisms found in LD. Thus, the
EcoRV and FokI polymorphisms were excluded from the
study, according to the LD findings. Permutation and raw
P values are presented in Table 4. A significant difference
in haplotype frequencies between melanoma cases and
controls was observed for At, aT, and at haplotypes. The
at haplotype was significantly more frequent in melanoma
patients than in the control group, where haplotype at
was not detected (6 vs. 0%, raw P = 0.003, permutation
P = 0.003). The At haplotype was more prevalent in
melanoma patients (39 vs. 32%, raw P = 0.040, permutation P = 0.210), whereas the aT haplotype was more
prevalent in the control group (40 vs. 29%, raw P = 0.030,
permutation P = 0.095).
The association between identified haplotypes and risk of
melanoma and clinicopathological characteristics was
estimated using the Thesias software. The aT haplotype
was associated with an increased risk of melanoma
(haplotype OR = 1.781, P = 0.006) in comparison with
the most frequent At haplotype (Table 4). A marginally
significant trend toward an association between the AT
haplotype and an increased risk of melanoma was found
(haplotype OR = 1.594, P = 0.052) (Table 4). No significant associations between any of the identified
haplotypes and clinicopathological characteristics were
observed (data not shown).

Genotype and allele distribution of vitamin D receptor gene polymorphisms in melanoma patients and the control group

Genes/SNPs
EcoRV/rs4516035

FokI/rs2228570

ApaI/rs7975232

TaqI/rs731236

Genotype

Cases (N)

Controls (N)

Allele

Cases (N)

Controls (N)

ee
Ee
EE
ff
Ff
FF
aa
Aa
AA
TT
Tt
tt

27
66
24
17
60
40
21
41
55
33
62
22

34
51
37
46
62
14
29
41
52
59
48
15

0.067

e
E

120
114

119
125

0.583

< 0.0001

f
F

94
140

154
90

< 0.0001

0.533

a
A

83
151

99
145

0.251

0.006

T
t

128
106

166
78

0.003

N, total number of cases/controls; SNP, single nucleotide polymorphism.

P < 0.05 is shown in bold.

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4 Melanoma Research 2014, Vol 00 No 00

Table 2

Vitamin D receptor gene polymorphisms association with the risk of melanoma

Genes/SNPs
EcoRV/rs4516035

FokI/rs2228570

ApaI/rs7975232

TaqI/rs731236

Genotype

Cases [N (%)]

Controls [N (%)]

ee
Ee
EE
Dominantb
Recessivec
Log-additive
ff
Ff
FF
Dominantb
Recessivec
Log-additive
aa
Aa
AA
Dominantb
Recessivec
Log-additive
TT
Tt
tt
Dominantb
Recessivec
Log-additive

27 (23)
66 (56)
24 (21)
90 (77)
93 (79)

17 (15)
60 (51)
40 (34)
100 (85)
77 (66)

21 (18)
41 (35)
55 (47)
96 (82)
62 (53)

33 (28)
62 (53)
22 (19)
84 (72)
95 (81)

34 (28)
51 (42)
37 (30)
88 (72)
85 (70)

46 (38)
62 (51)
14 (11)
76 (62)
108 (89)

29 (24)
41 (34)
52 (43)
93 (76)
70 (57)

59 (48)
48 (39)
15 (13)
63 (52)
107 (88)

Adjusted ORa (95% CI)

1.000
1.745
1.058
1.463
0.654
1.020
1.000
2.860
9.033
4.015
3.986
2.920
1.000
1.573
2.118
1.868
1.537
0.700
1.000
2.323
3.558
2.616
2.293
1.970

(0.8943.409)
(0.4332.524)
(0.7672.793)
(0.3441.245)
(0.6901.500)
(1.3975.855)
(3.49623.339)
(1.9838.128)
(1.9298.237)
(1.8504.590)
(0.7143.464)
(0.9654.650)
(0.9163.813)
(0.8712.714)
(0.4901.020)
(1.2194.427)
(1.5028.428)
(1.4334.775)
(1.0644.493)
(1.3002.970)

P
Reference
0.103
0.899
0.248
0.196
0.930
Reference
0.004
< 0.0001
< 0.0001
< 0.0001
< 0.0001
Reference
0.261
0.061
0.086
0.138
0.061
Reference
0.010
0.004
0.002
0.034
0.001

CI, confidence interval; N, total number of cases/controls; SNP, single nucleotide polymorphism; OR, odds ratio.
Adjusted for sex and age.
Heterozygous and mutated genotypes combined versus the wild-type genotype.
c
Mutated genotype versus heterozygous and the wild-type genotype combined.
P < 0.05 is shown in bold.
a

and serum levels of 25-hydroxyvitamin D (Supplementary

Table 3).

Table 3 Univariate analysis of clinicopathological variables of

melanoma patients according to the Cox regression model
95% CI
Variables
Sex
Age
Melanoma type
Ulceration
Breslow thickness
pT stage
Nodal status
Clinical stage at diagnosis
EcoRV rs4516035
FokI rs2228570
ApaI rs7975232
TaqI rs731236

HR

Lower

Upper

0.699
0.973
0.241
1.175
1.206
2.655
2.673
2.607
0.781
0.827
1.970
0.871

0.156
0.927
0.054
0.283
1.029
1.024
1.335
1.058
0.257
0.277
0.596
0.288

3.125
1.022
1.077
4.879
1.414
6.884
5.353
6.427
2.373
2.475
6.513
2.630

0.639
0.281
0.062
0.825
0.021
0.045
0.006
0.037
0.662
0.735
0.266
0.806

CI, confidence interval; HR, hazard ratio.

P < 0.05 is shown in bold.

25-Hydroxyvitamin D serum level

The 25-hydroxyvitamin D serum level was measured in

60 melanoma patients 36 months from diagnosis during
the winter months. Of melanoma patients, 31% had 25hydroxyvitamin D levels less than 25 nmol/l, 60% in a
range of 2650 nmol/l, whereas in only 9% of patients
concentrations were higher than 50 nmol/l. Thus, a serum
level of 25-hydroxyvitamin D was evaluated as a
dichotomous variable and classified into two categories:
less than 25 nmol/l and at least 25 nmol/l. There was no
association between 25-hydroxyvitamin D serum level
and VDR gene polymorphisms (Supplementary Table 2).
The absence of an association was observed for the
clinicopathological characteristics of melanoma patients

Discussion
Ultraviolet radiation plays an important role in melanomagenesis and, at the same time, it is necessary for the
skin vitamin D synthesis, the main source of vitamin D in
humans. The UVR action spectrum is almost the same for
direct DNA damage and vitamin D synthesis it belongs
to the UVB spectrum. Hence, there is a complex
relationship between UVR exposure, vitamin D levels,
VDR polymorphisms, and development of melanoma.
This could explain the variable association of these
factors in different populations from different latitudes,
and so far, these have been reported for the UK, Spain,
USA, and Denmark. In this study, we have explored these
associations, particularly VDR polymorphisms and vitamin
D levels, in samples in Serbia, representing the population of southeast Europe.
In our study, significant differences in both genotype
distribution and allele frequency of FokI and TaqI
VDR gene polymorphisms between the cases and control
groups indicate their potential role in melanoma susceptibility. The results of the current study indicate that
heterozygous and mutated genotypes, as well as the
variant allele carriers of FokI polymorphisms, could be
associated with an increased risk of melanoma in
comparison with the wild-type genotype. Thus, it could
be assumed that wild-type FokI could play a protective
role in melanoma susceptibility. Our findings are in

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VDR gene polymorphisms in melanoma Zeljic et al.

Fig. 1

18

8
9

78

Block 1 (0 kb)
3
4
2

0
0

18
1

rs731236

rs7975232

rs2228570

Block 1 (0 kb)
3
4

rs4516035

rs731236

rs2228570

rs7975232

rs4516035

(a)

24
2

16

16
1

59

Block 1 (0 kb)
3
4
2

12
5

rs731236

rs7975232

rs2228570

Block 1 (0 kb)
3
4

rs4516035

rs731236

rs2228570

rs7975232

rs4516035

(b)

17
0

11

19
15

rs731236

rs7975232

1
0

12
8

rs2228570

Block 1 (0 kb)
3
4

rs4516035

rs731236

rs2228570

rs7975232

rs4516035

(c)

Block 1 (0 kb)
3
4
1

32
1

Pairwise linkage disequilibrium plots of analyzed polymorphisms in the vitamin D receptor gene for (a) cases and the control group taken together,
(b) cases group, (c) control group. D0 and r2 values are presented in percentages. The red-shaded boxes correspond to the paired D0 values between
the single nucleotide polymorphisms (SNPs). Shades of pink: LOD > 2, D0 > 1; red: LOD > 2, D0 = 1; white: LOD < 2, D0 < 1. The gray-shaded boxes
correspond to the paired r2 between the SNPs. White: r2 = 0; shades of gray: 0 < r2 < 1; black: r2 = 1. LOD, logarithm of odds.

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6 Melanoma Research 2014, Vol 00 No 00

Table 4 Haplotype frequencies and haplotype odds ratio analysis for vitamin D receptor gene polymorphisms, compared with the most
common haplotype
N (%)
Haplotypes
At
aT
AT
at

Total
85
84
63
7

(36)
(35)
(26)
(3)

Association P value

Cases

Controls

46
35
29
7

39
50
34
0

(39)
(29)
(25)
(6)

(32)
(40)
(28)
(0)

Raw

Permutation

HOR (95% CI)

0.040
0.030
0.336
0.003

0.210
0.095
0.769
0.003

Reference
1.781 (1.1762.698)
1.594 (0.9962.551)
0.362 (0.00344.178)

0.006
0.052
0.678

N, total number of cases/controls; HOR, haplotype odds ratio; CI, confidence interval.
P < 0.05 is shown in bold.

agreement with the results from previous studies on

melanoma [2326] and basal cell carcinoma [27],
although contrasting the results for oral cancer patients
from the Serbian population [19]. It is known that the
mutated form of FokI results in the synthesis of a shorter
receptor form with higher transactivation capacity as a
transcription factor [15]. Different findings of other
studies examining various cancer types could be explained by the gene and cell type-specific transactivation
capacity of the FokI mutated form [15].

previous findings in melanoma [2,25,29] and oral carcinoma

[19]. However, a previous study has reported that the EcoRV
mutated genotype is associated with a decreased risk of
cutaneous melanoma compared with the wild-type form [30].
In another study, the wild-type allele of EcoRV SNP was
associated with an increased susceptibility to melanoma, risk
for metastasis, and poor patient outcome [31]. Conflicting
data from the different studies warrant further analysis to
elucidate the role of the EcoRV polymorphism in melanoma
susceptibility.

Variant allele carriers of TaqI polymorphism were

associated with an increased risk of melanoma, which is
consistent with findings from other study [3]. In contrast,
a recent study has reported no association of the TaqI
polymorphism with the risk of melanoma [28]. The
inconsistency between studies could be explained by
population differences as well as study group size. Our
data suggest that the wild-type genotype might protect
carriers against melanoma. It is known that the TaqI
polymorphism does not alter the amino acid sequences of
the VDR protein [3]. Thus, the exact mechanism of how
the TaqI polymorphism could influence the risk of
melanoma remains unclear. It could be assumed that
the mutated variant is itself functional or is in LD with
other functionally relevant polymorphisms in the VDR
gene. Further investigations on the functional effects of
VDR polymorphisms on the risk of melanoma are
warranted.

Studies on the association between VDR gene polymorphisms and the risk of melanoma are not consistent in
the literature. These discrepancies may be because of
differences in serum vitamin D levels, sun exposure,
population differences, and sample size [3]. A study
carried out in a Danish population showed that elevated
plasma calcidiol level was associated with an increased
risk of nonmelanoma and melanoma cancer [14]. In
contrast, higher vitamin D serum levels were associated
with lower Breslow thickness and better survival of
melanoma patients in a UK study [12]. In the first study
from a country with high insolation (Spain), suboptimal
vitamin D levels were also found in melanoma patients at
diagnosis, but the levels were higher than those reported
in the UK study. In our study, the vitamin D levels in
winter months were from 20 to 75 nmol/l in 68.6% of
patients and less than 20 nmol/l in 31.4% of patients
(median age 48.5 years). Low vitamin D levels were also
found in postmenopausal women in this region in 88.4%
of patients [32]. Apart from the possible insufficient sun
exposure, these data also indicate a probable insufficient
vitamin D intake in our population, and this was
reported previously for the region of central and eastern
Europe [33], possibly linked to socioeconomic factors,
compared with western European countries [33]. This
further emphasizes the necessity of identifying melanoma
patients with suboptimal vitamin D levels, and advising
them on vitamin D supplementation as after the
diagnosis of melanoma, rigorous sun protection is advised,
and therefore, vitamin D status could deteriorate further.

VDR ApaI and TaqI polymorphisms were found in LD in

our study group, which is in agreement with the literature
data [15]. Among the haplotypes identified, the aT
haplotype was significantly associated with an increased
risk of melanoma. The ApaI polymorphism was not
associated with the risk of melanoma itself, whereas in
the haplotype block with the T allele of TaqI SNP,
association with increased susceptibility to melanoma was
observed. These findings indicate that genetic association
studies should include haplotype-based analysis. In
addition, it has been shown that ApaI and TaqI
polymorphisms could influence VDR mRNA stability [15],
which could be a possible explanation for the haplotype
association observed in the current study.
The lack of association of the EcoRV polymorphism with the
risk of melanoma observed in our research is consistent with

There was no association between vitamin D serum level

and VDR gene polymorphisms analyzed nor with clinicopathological characteristics in melanoma patients,
which is consistent with findings from other studies.
Because of the small number of patients, the previously

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VDR gene polymorphisms in melanoma Zeljic et al.

described interaction of VDR polymorphism and vitamin

D level could not be explored [12].

15

The rapid increase in the incidence of melanoma and its

high mortality requires comprehensive research on the
risk factors associated with melanoma. Our results
suggest an association between FokI and TaqI polymorphisms in the VDR gene and the risk of melanoma,
implicating their potential application as molecular
markers for the management of patients at high risk for
the development of this cancer and their increased
surveillance. Further analysis on vitamin D intake and
serum calcidiol levels of melanoma patients, as well as
genegene and geneenvironment interactions, would
provide additional information on the importance and role
of the VDR gene in melanoma susceptibility.

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Acknowledgements
Conflicts of interest

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There are no conflicts of interest.

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