Beruflich Dokumente
Kultur Dokumente
Abstract
Prostate cancer is the second most common cancer in men worldwide. Although the aetiology of this disease
remains largely unclear, several lines of evidence suggest that oxidative stress plays a role in prostate carcinogenesis. The antioxidant enzyme glutathione peroxidase 1 (GPX1) is part of the enzymatic antioxidant defence,
preventing oxidative damage to DNA, proteins and lipids by detoxifying hydrogen and lipid peroxides that may
contribute to prostate cancer development. Some studies indicate an association between GPX1 Pro198Leu
polymorphism and an increased risk of cancer. The purpose of the present study was to determine the possible
association of GPX1 Pro198Leu polymorphism and erythrocyte GPX activity with the risk of developing
prostate cancer and to clarify whether erythrocyte GPX activity levels were correlated with the GPX1
Pro198Leu genotype in the Turkish population. The GPX1 Pro198Leu genotype was determined in 33 prostate
cancer patients and 91 control individuals. As evident from our results, there was no difference between
genotype and/or allele frequencies in prostate cancer patients and controls. No significant difference was found
in GPX1 genotype or allele frequency between aggressive and non-aggressive prostate cancer patients. It can
be suggested with these findings that individual susceptibility of prostate cancer may be modulated by GPX1
polymorphism, but it needs further studies.
Keywords
prostate cancer; glutathione peroxidase 1; genetic polymorphism; oxidative stress
Introduction
Prostate cancer continues to be a major age-related
malignancy with most incidences occurring between
54 and 75 years and rapid onset after 45 years. Evidence from epidemiological, experimental and clinical studies suggests that prostate cancer cells are
exposed to an increased oxidative stress.1,2 Reactive
oxygen species (ROS), most notably the hydroxyl
radicals, generated endogenously by cellular metabolism are known to cause oxidative DNA damage that
has been implicated in prostate carcinogenesis.3
The antioxidant enzyme glutathione peroxidase 1
(GPX1) is part of the enzymatic antioxidant defence,
preventing oxidative damage to DNA, proteins and
lipids by detoxifying hydrogen and lipid peroxides.4,5
The cytosolic form of GPX1 belongs to a family of
selenium-dependent peroxidases that include cytosolic GPX,6 plasma-based GPX37 and phospholipids
hydroperoxidase GPX4.8 GPX1 knockout mice have
Corresponding author:
Ahmet Aydn, Department of Toxicology, Yeditepe University, 26
Agustos Yerlesimi, Kayisdagi, Istanbul 34755, Turkey
Email: ahmetaydin30@hotmail.com
Erdem O et al.
25
26
reaction mixture containing 1 mmol/L Na2EDTA, 2 Table 1. Comparison of cases and controls by selected
mmol/L reduced glutathione, 0.2 mmol/L NADPH, demographic and clinical variables
4 mmol/L sodium azide and 1000 U glutathione Characteristic
Cases
Controls
p Valuea
reductase in 50 mmol/L TRIS buffer (pH 7.6) was
Age (years, mean + SD)
67.52 + 9.31 64.57 + 8.72
0.115
prepared. A total of 20 ml of erythrocyte lysate and Smoking status (n, %)
0.088
980 ml of the reaction mixture were mixed and incuNever
11 (33.3)
29 (31.9)
Current
4 (12.1)
19 (20.9)
bated for 5 min at 37 C. The reaction was initiated
18 (54.5)
43 (47.3)
by adding 8.8 mmol/L hydrogen peroxide and the PSAFormer
(ng/ml)
0.003100
0.00310.31
decrease of absorbance was recorded at 340 nm for Free PSA (ng/ml)
0.16 + 0.25 0.65 + 0.67
0.004
Risk level (n, %)
3 min. GPX activity is expressed in U/ml.
Non-aggressive disease
Aggressive disease
Statistical analysis
Demographic information stratified by casecontrol
status was tabulated as a mean + standard deviation
for continuous variables and a number (and percentage) for categorical variables. Pearsons w2 test was
used to assess group differences on categorical
variables and a two-sample t test was used to assess
group differences for continuous variables. Comparison of the erythrocyte GPX activity between cases
and controls was carried out by a two-sample t test.
A KruskalWallis non-parametric analysis of variance (ANOVA) was used to assess whether mean
concentration of erythrocyte GPX activities varied
by genotype among the controls. Allele and genotype
frequencies of cases and controls were compared with
values predicted by HardyWeinberg equilibrium
using the w2 test. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated to evaluate
the associations between GPX1 genotypes and prostate risk. Calculation for the casecontrol study was performed using the DeFinetti computer program, http://
ihg.gsf.de/cgi-bin/hw/hwa1.pl. In all cases, p values
0.05 were considered statistically significant.
Additionally, study subjects were stratified according to age (based on age at diagnosis for cases or age at
selection for controls) to evaluate the relationship of
the GPX1 polymorphism with early-onset prostate
cancer.
The association of the GPX1 polymorphism and
disease status was studied with refitted models for
non-aggressive and aggressive prostate cancer, respectively. Men diagnosed with high-grade cancer
(Gleason score of 710) were categorized as having
aggressive disease; those diagnosed with low-grade
prostate cancer (Gleason score 26) were categorized
as having non-aggressive disease.
For all analysis, we first examined the association
of GPX1 Pro/Leu and Leu/Leu genotype, using Pro/
Pro genotype as a reference. Next, as a result of the
10 (30.3)
23 (69.7)
Cases
Controls
p Valuea
33
7.14 + 1.20
91
8.17 + 1.56
0.0007
lack of complete information on the functional significance of GPX1 Pro198Leu polymorphism, we analysed the data under the assumption of both
dominant (grouping heterozygous with homozygous
rare allele) and recessive models (grouping heterozygous with wild type).
Results
Subject characteristics
Table 1 shows a casecontrol comparison of selected
baseline subject characteristics. As expected, free
prostate specific antigen (PSA) levels were significantly
different in cases compared with controls. Age and
smoking status were not different in prostate cancer
cases compared to controls.
Erdem O et al.
27
Cases (n, %)
Controls (n, %)
OR
95% CI
(33.3)
(51.5)
(15.2)
(84.8)
(15.2)
(33.3)
(66.7)
40 (44.0)
41 (45.1)
10 (11.0)
81 (89.1)
10 (11.0)
40 (44.0)
51 (56.1)
1.00
0.62
1.82
1.00
0.8
1.00
1.57
Reference
0.221.76
0.516.44
Reference
0.361.76
Reference
0.683.61
39 (59.1)
27 (40.9)
121 (66.5)
61 (33.5)
1.00
1.16
Reference
0.592.28
11
17
5
28
5
11
22
p Value
0.367
0.350
0.572
0.288
0.660
Discussion
In the present study, we found there was no difference
between genotype and/or allele frequencies in prostate cancer cases and controls. Few studies have
investigated the relationship of GPX1 Pro198Leu and
prostate cancer risk.2,25 Choi et al.25 failed to find
associations between GPX1 Pro198Leu polymorphism and prostate cancer risk among men with a history
of smoking and/or asbestos exposure. Further analyses stratified by factors related to environmental
oxidative stress did not modify associations.25 On the
contrary, Arsova-Sarafinovska et al.2 found an overall
protective effect of the variant Leu allele of the GPX1
polymorphism on the prostate cancer risk.
Furthermore, in our study we found lower erythrocyte GPX activity in the cancer group than in the healthy
controls. These data confirmed our results obtained both
in Macedonian and Turkish populations. It was also
published in previous studies in which we reported that
lower GPx activity was associated with prostate
cancer.2,26 There are variable reports on the activity of
this enzyme in prostate cancer. Jung et al.27 found
no differences in the antioxidant enzymatic activities
of prostatic epithelial cell cultures between benign
and malign tissue. In other studies, malignant epithelial
cells in prostatic adenocarcinoma have been found
28
Table 4. GPX1 genotype and allele frequencies and ORs (95% CI) in cases and controls stratified by age at diagnosis (for
cases) and age at selection (for controls)
GPX1 genotype
Age 65 years
Genotype frequencies
Pro/Pro
Pro/Leu
Leu/Leu
Pro/Pro and Pro/Leu
Leu/Leu
Pro/Pro
Pro/Leu and Leu/Leu
Allele frequencies
Pro
Leu
Age 65 years
Genotype frequencies
Pro/Pro
Pro/Leu
Leu/Leu
Pro/Pro and Pro/Leu
Leu/Leu
Pro/Pro
Pro/Leu and Leu/Leu
Allele frequencies
Pro
Leu
Cases (n, %)
Controls (n, %)
OR
95% CI
8 (57.1)
4 (28.6)
2 (14.3)
12 (85.7)
2 (14.3)
8 (57.1)
6 (42.9)
20 (37.0)
26 (48.1)
8 (14.8)
46 (85.1)
8 (14.8)
20 (37.0)
34 (62.9)
1.00
0.39
0.63
1.00
0.958
1.00
0.44
Reference
0.11.46
0.113.6
Reference
0.185.11
Reference
0.131.46
20 (71.4)
8 (28.6)
66 (61.1)
42 (38.9)
1.00
0.63
Reference
0.251.56
3 (15.8)
13 (68.4)
3 (15.8)
16 (84.2)
3 (15.8)
3 (15.8)
16 (84.2)
19 (52.8)
15 (41.7)
2 (5.6)
34 (94.5)
2 (5.6)
19 (52.8)
17 (97.3)
1.00
5.49
9.5
1.00
0.34
1.00
5.96
Reference
1.3222.85
4.0982.73
Reference
0.091.3
Reference
1.4824.08
19 (50.0)
19 (50.0)
53 (73.6)
19 (26.4)
1.00
2.79
Reference
1.226.36
p Value
0.152
0.597
0.960
0.173
0.313
0.014
0.024
0.103
0.008
0.013
Table 5. GPX1 genotype and allele frequencies and ORs (%95 CI) in aggressive and non-aggressive prostate cancer
GPX1 genotype
Genotype frequencies
Pro/Pro
Pro/Leu
Leu/Leu
Pro/Pro and Pro/Leu
Leu/Leu
Pro/Pro
Pro/Leu and Leu/Leu
Allele frequencies
Pro
Leu
Non-aggressive prostate
cancerb (n, %)
OR
95% CI
(34.8)
(47.8)
(17.4)
(82.6)
(17.4)
(34.8)
(65.2)
3 (30.0)
6 (60.0)
1 (10.0)
9 (90.0)
1 (10.0)
3 (30.0)
7 (70.0)
1.00
0.69
1.5
1.00
1.26
1.00
0.8
Reference
0.133.6
0.1219.44
Reference
0.275.93
Reference
0.163.99
27 (58.7)
19 (41.3)
12 (60.0)
8 (40.0)
1.00
1.06
Reference
0.363.08
Aggressive prostate
cancera (n, %)
8
11
4
19
4
8
15
p Value
0.657
0.756
0.767
0.789
0.921
Erdem O et al.
29
Table 6. Erythrocyte GPX activity by the Pro198Leu polymorphism in the GPX1 gene in cases and controls
Cases (n 33)
GPX1 genotype
Pro/Pro
Pro/Leu
Leu/Leu
pb
7.42 + 1.50
7.05 + 0.99
6.81 + 1.24
0.6008
Controls (n 91)
n (%)
n (%)
11 (33.3)
17 (51.5)
5 (15.2)
8.37 + 1.36
7.88 + 1.61
8.60 + 2.00
0.254
40 (44.0)
41 (45.1)
10 (11.0)
GPX1: glutathione peroxidase 1, GPX: glutathione peroxidase, SD: standard deviation, ANOVA: analysis of variance.
a
Values are indicated by mean + SD.
b
p from KruskalWallis non-parametric ANOVA test for difference in GPX activity by GPX1 genotype.
30
References
1. Khandrika L, Kumar B, Koul S, et al. Oxidative stress
in prostate cancer. Cancer Lett 2009; 282: 125136.
2. Arsova-Sarafinovska Z, Matevska N, Eken A, et al.
Glutathione peroxidase 1 (GPX1) genetic polymorphism, erythrocyte GPx activity, and prostate cancer risk.
Int Urol Nephrol 2009; 41: 6370.
3. Trzeciak AR, Nyaga SG, Jaruga P, et al. Cellular repair
of oxidatively induced DNA base lesions is defective
in prostate cancer cell lines, PC-3 and DU-145. Carcinogenesis 2007; 28: 13591370.
4. Ravn-Haren G, Olsen A, Tjonneland A, et al. Associations between GPX1 Pro198Leu polymorphism, erythrocyte GPX activity, alcohol consumption and
breast cancer risk in a prospective cohort study. Carcinogenesis 2006; 27: 820825.
5. Arthur JR. The glutathione peroxidases. Cell Moll Life
Sci 2000; 57: 18251835.
6. Chu FF, Doroshow JH, and Esworthy RS. Expression,
characterization, and tissue distribution of a new cellular selenium-dependent glutathione peroxidase,
GSHPx-GI. J Biol Chem 1993; 268: 25712576.
7. Takahashi K, Akasaka M, Yamamoto Y, et al. Primary
structure of human plasma glutathione peroxidase
deduced from cDNA sequences. J Biochem 1990;
108: 145148.
Erdem O et al.
21.
22.
23.
24.
25.
26.
31
27.
28.
29.
30.
31.
32.
Copyright of Human & Experimental Toxicology is the property of Sage Publications, Ltd. and its content may
not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written
permission. However, users may print, download, or email articles for individual use.