Detection of Chromosomal Translocation in Prostate Cancer and Benign Prostatic Hyperplasia by Fluorescence in situ Hybridization (FISH)

1
Detection of Chromosomal Translocation in
Prostate Cancer and Benign Prostatic Hyperplasia
by Fluorescence in situ Hybridization (FISH)
Cebulska-Wasilewska A.1,2, Miszczyk J.1,
Dobrowolska B.3, Dobrowolski Z.3

1 Environmental and Radiation Biology Department,
The H. Niewodniczanski Institute of Nuclear
Physics PAN, Cracow, Poland, 2 Epidemiology and
Preventive Medicine Departmen CM UJ, Poland,
3Urology Department and Clinic CM UJ, Poland

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Prostate cancer epidemiology
Benign Prostatic Hyperplasia BPH and Prostate
Cancer PC are the most common males diseases.
Prostate cancer incidence varies widely
between ethnic populations, countries and
increases sharply with older age.
In Poland prostate cancer is the third most
common malignant cancer in males.

3
Prostate cancer risk is strongly influenced by
GENETIC FACTORS
FAMILIAL HISTORY
EPIGENETIC FACTORS
gt genetic susceptibility
gt lifestyle, smoking
gt genes involved in familial prostate cancer
(for example HPC1 on chromosome 1)
gt ethnic origin
gt mutation
gt dietary factors (vitamin D, fats,)
gt gene expression
gt androgens
4







Becouse...

• gt genetic alteration on multiple chromosomes
  including especially chromosome 1,
• gt many susceptibility loci have been reported at
  this chromosome,
• gt many types of cancers are associated with
  specific types of chromosomal aberrations

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Aim of study
Compare the vulnerability to the induction in
chromosome 1 translocation in lymphocytes from
prostate cancer with that from benign prostatic
hyperplasia.
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Materials and methods
Investigated groups
Standard cytogenetic procedure
Fig. 1.
BPH 27 persons from the control group with
benign prostatic hyperplasia. (average age 68.9
8.3)
PCP 30 prostate cancer patients. (average
age 62.45.3)
Challenging dose
In the laboratory tubes with blood were
irradited with X-rays doses of 2 Gy.
Culture were set up according to standard
cytogetic procedure, then were harvested and
followed by fixation procedure.
Fluorescent in situ hybridisation (FISH)
Biotin-labeled whole chromosome probes specific
to chromosome 1 (Star Fish Cambio, UK).
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Materials and methods
General FISH protocol
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.
The slides were examined at 1000x magnification
of the epifluoescence microscope (Nicon Eclipse
E400).
Donors were examined for presence in their in
peripheral blood lymphocytes of chromosome
translocations according to the criteria of
Protocol for Aberration Identification and
Nomenclature-PAINT 1.
1 Tucker J.D. et al. A proposed system for
scoring structural aberrations detected by
chromosome painting. Cytog. Cell Genet. (1995),
211-221.
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Materials and methods
2 types of parameters were used to describe the
extent of chromosomal damage
t frequency of chromosome 1 translocation
FG/100 genomic frequency of chromosome 1
translocation
FGFg/2.05fp(1-fp) 2.
FG - the total genomic translation frequency
Fg the translocation frequency measured by FISH
after painting
Fp the fraction of th genome represented the
painted chromosome, for chromosome 1 0.084
fraction of the genome 8.4)
2 Lucas J.N., Sachs R.K. Using three-color
chromosomepainting to test chromome aberration
models. Proc. Natl. Sci90, 1484-14
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Results
Sig. 1. X-rays effect on frequency of chromosome
1 translocation determined by FISH in peripheral
lymphocytes in patients with prostate cancer
PCP and benign prostate hyperplasia BPH.
t
T number of translocations/1000 cells
Number of translocations/1000 cells (t) was
significantly higher in patients with prostate
cancer (14.600.91) than in the control group
(10.24 1.10 plt0.01).
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Results
Sig. 2. Genomic frequency of chromosome 1
translocation for patients with prostate cancer
PCP and benign prostatic hyperplasia BPH.
FG/100
FG/100 - genomic frequency of translocation
Percentage of FG/100 was significantly higher in
patients with prostate cancer (0.550.03) that
obtained for the reference group (0.38 0.04,
plt0.01).
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We want to study correlation between occur
cancer in family and frequency of chromosome 1
translocation.
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Results
Tab. 1. Correlations for patients with prostate
cancer between age of donors, existing cancer in
the closely related members of family and t.
t
Age R 0.50
Age plt .001
CiF R 0.11
CiF plt .001
CiF reported cancer in the immediate family
t number of translocations/1000 cells
R correlation coefficient
High and significant correlation between age of
donors and frequency of chromosome 1
translocation was observed (0.50 plt0.001).
Furthermore, there was also correlation between
frequency of chromosome 1 translocation observed
in patients who had reported other cancers in
family.
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Conclusions
1. These studies, although preliminary, are
suggesting that frequency of translocation
detected in the response to challenging treatment
might be used as predictor of susceptibility for
prostate cancer patients.
2. Our results might confirm hypothesis that
exist an association between predisposition to
genetic instability chromosome 1 and hereditary
or familial conditioning of prostate cancer.
However, more studies are necessary of other
factors which could affect genomic frequency of
translocations such as life style, diet or
genetic polymorphism.