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
1Environmental and Radiation Biology Department, The H. Niewodniczański Institute of
Nuclear Physics PAN, Cracow, Poland, 2 Epidemiology and Preventive Medicine
Departmen CM UJ, Poland, 3Urology Department and Clinic CM UJ, Poland
Prostate cancer epidemiology
Benign Prostatic Hyperplasia [BPH] and
Prostate Cancer [PC]
are the most common males diseases.
Prostate cancer incidence
In Poland prostate varies widely between:
cancer is the third ethnic populations, countries
most common and increases sharply with
malignant cancer in older age.
Prostate cancer risk is strongly influenced by:
GENETIC FAMILIAL EPIGENETIC
FACTORS HISTORY FACTORS
> genetic > genes involved in > lifestyle, smoking
susceptibility familial prostate cancer > ethnic origin
> mutation (for example: HPC1 on
chromosome 1) > dietary factors
> gene (vitamin D, fats,)
> genetic alteration on multiple chromosomes including
especially chromosome 1,
> many susceptibility loci have been reported at this
> many types of cancers are associated with specific
types of chromosomal aberrations
Aim of study
Compare the vulnerability to the induction in
chromosome 1 translocation in lymphocytes
from prostate cancer with that from benign
Materials and methods
Investigated groups Fig. 1. Standard cytogenetic procedure
antibiotics, PHA colcemid
[BPH] – 27 persons from the X-rays Incubation at
irradiation 37ºCfor 72 hours
control group with benign
(average age 68.9 ± 8.3) Donor’s peripheral 80% RPMI 1640,
blood samples 20% fetal calf serum
[PCP] – 30 prostate cancer patients.
(average age 62.4±5.3) KC
Challenging dose Fixation methanol/acetic acid
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
Materials and methods
Fig. 2. General FISH protocol
probe Hybridization 37ºC,
Place probe Fig. 4.
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 .
 Tucker J.D. et al. A proposed system for scoring structural aberrations detected by chromosome painting.
Cytog. Cell Genet. (1995), 211-221.
Materials and methods
2 types of parameters were
used to describe the extent of
t – frequency of chromosome 1 FG/100 – genomic frequency of
translocation chromosome 1 translocation
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%)
 Lucas J.N., Sachs R.K. Using three-color chromosomepainting to test chromome aberration models. Proc. Natl.
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 – number of translocations/1000 cells
Number of translocations/1000 cells (t) was significantly higher
in patients with prostate cancer (14.60±0.91) than in the control
group (10.24 ±1.10; p<0.01).
Sig. 2. Genomic frequency of chromosome 1 translocation for patients with
prostate cancer [PCP] and benign prostatic hyperplasia [BPH].
FG/100 - genomic frequency of translocation
Percentage of FG/100 was significantly higher in patients with
prostate cancer (0.55±0.03) that obtained for the reference group
(0.38 ±0.04, p<0.01).
We want to study correlation between
occur cancer in family and frequency of
chromosome 1 translocation.
Tab. 1. Correlations for patients with prostate cancer between age of donors,
existing cancer in the closely related members of family and t.
CiF – reported cancer in the immediate family t
t – number of translocations/1000 cells Age R 0.50
R – correlation coefficient
CiF R 0.11
High and significant correlation between age of donors and
frequency of chromosome 1 translocation was observed (0.50;
Furthermore, there was also correlation between frequency of
chromosome 1 translocation observed in patients who had reported
other cancers in family.
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.