Post on 07-May-2015
description
transcript
STUDY OF THE ROLE OF
ESTROGEN RECEPTOR ALPHA GENE POLYMORPHISM
IN PROSTATE CANCER AMONG EGYPTIANS
By
Ola Hussein Elgaddar
Assistant lecturer of Chemical Pathology
Medical Research Institute
Alexandria University
Professor Dr. Thanaa Fathy Moghazy Professor of Clinical Pathology
Department of Chemical Pathology
Medical Research Institute
University of Alexandria
Professor Dr. Saad Mohammed Saad Professor of Urology
Department of Urology
Faculty of Medicine
University of Alexandria
Professor Dr. Amel Abd El-Fattah Kamel Professor of Chemical Pathology
Department of Chemical Pathology
Medical Research Institute
University of Alexandria
Dr. Moyassar Ahmad Zaki Assistant Professor of Chemical Pathology
Department of Chemical Pathology
Medical Research Institute
University of Alexandria
Types of Estrogen
Estrogen receptors:
Estrogens induce cellular changes
through binding to its specific receptors.
There are two described types of
estrogen receptors (ERs) in humans:
ER alpha (ERα)
ER beta (ERβ)
Comparison of the structures and homology between ERα and ERβ
Estrogen receptor genes:
ERα and ERβ are encoded by ESR1
and ESR2 genes, present on
chromosomes 6q25.1 and 14q23-24.1,
respectively.
ESR1 and ESR2 comprise eight exons
separated by seven intronic regions and
spans more than 140 kilo-bases and
approximately 40 kilobases, respectively
Genomic format and domain structure of human ERα and ERβ
Estrogens – ER signaling pathways:
Classified in two pathways; genomic
and non-genomic
A. Genomic (Classical) Pathway:
E2 binds to nuclear or cytoplasmic receptors.
E2 actions occur over the course of hours.
B. Non-Genomic Pathway:
E2 binds to ER located in or adjacent to the
plasma membrane
E2 actions occur within few minutes
ER gene variants:
The DNA variants (polymorphisms) are
common, occurring in more than 1% of the
population.
The most common type of polymorphisms is a
single nucleotide change in DNA sequence
and is referred to as single nucleotide
polymorphisms (SNPs).
SNPs may account for many well-
characterized phenotypes, including disease
susceptibility and resistance.
Another type of polymorphisms is the
tandemly repeated DNA sequences.
They represent variation in the number of
copies of a tandemly repeated motif at each
locus.
These repeats differ in length to form variable
number tandem repeat (VNTR)
VNTR
ERα gene polymorphism:
The most widely studied are:
T397C (PvuII: rs2234693)
A351G (XbaI: rs9340799)
Both named after the relevant restriction enzyme
Both lie in intron I in the ERα gene
They are in strong linkage disequilibrium
(TA)n variable number of tandem repeats
within the promoter region of the gene
Though both PvuII and XbaI polymorphisms
lie in intronic (apparently non-functioning) part
of the gene, they have been widely studied in
many diseases as they thought to be:
In linkage disequilibrium with causal
polymorphisms elsewhere in the ERα gene or
in an adjacent gene.
In linkage disequilibrium with the upstream
TA repeat polymorphism in the promoter
region of the ERα gene, having a significant
influence on transcriptional regulation
THE PROSTATE GLAND:
The prostate is the major accessory sex
gland of the male.
Its secreted fluid comprises 15% of the
ejaculate.
The gland has been the subject of much
study being susceptible to infection as well as
neoplastic transformation.
Carcinoma of the prostate: The 6th most common cancer in the world
and the 3rd in importance in men.
The prostate cancer is genetically
heterogeneous, with several genes having
different frequencies and penetrance.
Environmental factors may have a role in
inducing different genetic processes and
molecular pathways.
Heterogeneity of both environmental factors
and susceptibility genes could explain the
discrepancies of prostate cancer incidence
between populations.
The final diagnosis of prostate cancer is
done by histopathological examination of a
biopsy and it is graded by the Gleason system
(score).
Staging of the tumor is done using the tumor
node metastases (TNM) system, that had been
adopted by the American Joint Committee for
Cancer Staging.
Estrogens and their receptors in
cancer prostate: Prostate expresses both ER-α (mainly
stromal) and ER-β (mainly epithelial)
Evidence demonstrates a clear dichotomy
between ER-α and ER-β actions. The adverse
effects of estrogen, via ER-α, are specifically
related to the development of prostatic
proliferation and inflammation as well as
prostate cancer. In contrast, the beneficial
effects of estrogens reside with the activation of
ER-β, which appears to mediate the anti-
proliferative, anti-inflammatory and, potentially,
anti-carcinogenic effects of estrogen.
Estrogen receptor-α, coded by the ESR1
gene, has been examined in several studies for
the presence of polymorphic sites and their
relation to cancer prostate.
Among the most commonly studied
polymorphisms are the ESR1 T397C (PvuII) and
A351G (XbaI) restriction fragment length
polymorphism (RFLP) markers.
Results regarding the presence or absence of
an association between these two
polymorphisms and the presence of cancer
prostate are controversial among different
populations.
The present study aimed at
studying the role of T397C and
A351G polymorphisms of
estrogen receptor alpha gene in
the occurrence of prostate
cancer among Egyptians.
100 Egyptian males
Group III
Cancer prostate
N = 45
Group II
BPH
N = 20
Group I
Controls
N = 35
To all studied subjects the
following was done:
I) Full clinical examination
II) Laboratory investigations: Preliminary tests in serum.
(Creatinine, ALT, ALP, ACP; total & prostatic)
Serum tPSA
Serum Estradiol (E2)
Molecular studies for the detection of T397C
(PvuII) and A351G (XbaI) polymorphisms in
estrogen receptor α gene
Molecular study:
1) DNA extraction
2) PCR
3) RFLP
10
0 b
p la
dd
er D
NA
mar
ker
346 bp PCR
product
10
0 b
p la
dd
er
DN
A m
arke
r
ERα T397C (PvuII) genotype distribution, according to the
Hardy-Weinberg equilibrium among the studied groups
0
5
10
15
20
25
30
CC CT TT CC CT TT CC CT TT
Control group BPH group Cancer prostate group
Observed frequency
Expected frequency
Chi-sq: 0.031
P-value: 0.985
Chi-sq: 0.312
P-value: 0.876
Chi-sq: 2.212
P-value: 0.331
ERα T397C (PvuII) genotype distribution
among the studied groups
TT CT CC TT CT CC TT CT CC
0
10
20
30
40
50
60
70
80
90
100
Control group BPH group Cancer prostate group
22.9
30 26.7
51.4 55
60
25.7
15 13.3
%
TT
CT
CC
48.60% 51.40%
T
C
ERα T397C (PvuII) allele frequencies in
the control group
57.50% 42.50%
T
C
ERα T397C (PvuII) allele frequencies in
the BPH group
56.70% 43.30%
T
C
ERα T397C (PvuII) allele frequencies in
the cancer prostate group
ERα T397C (PvuII) allele frequencies in
the studied groups
T C T C T C
0
10
20
30
40
50
60
70
80
90
100
Control group BPH group Cancer prostate group
48.6
57.5 56.7
51.4
42.5 43.3
%
T
C
ERα A351G (XbaI) genotype distribution, according to the
Hardy-Weinberg equilibrium among the studied groups
0
5
10
15
20
25
30
AA / AG GG AA / AG GG AA /AG GG
Control group BPH group Cancer prostate group
Observed frequency
Expected frequency
Chi-sq: 0.141
P-value: 0.707
Chi-sq: 0.317
P-value: 0.573
Chi-sq: 0.915
P-value: 0.339
ERα A351G (XbaI) genotype distribution
among the studied groups
AA AG GG AA AG GG AA AG GG
0
10
20
30
40
50
60
70
80
90
100
Control group BPH group Cancer prostate group
0 0 0
34.3
50 53.3
65.7
50 46.7
%
AA
AG
GG
17.10%
82.90%
A
G
ERα A351G (XbaI) allele frequencies in
the control group
ERα A351G (XbaI) allele frequencies in
the BPH group
25 %
75% A G
ERα A351G (XbaI) allele frequencies in
the cancer prostate group
26.70%
73.30% A
G
ERα A351G (XbaI) allele frequencies in
the studied groups
A G A G A G
0
10
20
30
40
50
60
70
80
90
100
Control group BPH group Cancer prostate group
17.1
25 26.7
82.9
75 73.3
%
A
G
1) No statistically significant difference
could be detected in both T397C (PvuII)
and A351G (XbaI) polymorphisms of the
ERα gene, between the three studied
groups of Egyptians; namely cancer
prostate patients, benign prostatic
hyperplasia patients and healthy controls.
Thus, the development of cancer prostate
among Egyptian males cannot be
attributed to these two polymorphisms.
2) No association could be found
between the presence of variant alleles
(C allele of PvuII and G allele of XbaI
polymorphisms of ERα gene), and the
development of cancer prostate among
the studied sample of Egyptian male.
Larger population based studies are
needed to assess the association
between T397C (PvuII) and A351G
(XbaI) polymorphisms of the ERα
gene, and the occurrence of cancer
prostate in Egyptians.
Other polymorphisms may be
studied in ER α, in relation to the
development of cancer prostate,
such as TA repeats in the promoter
region.