2008 Barcelona - European Society of Human Genetics

Cancer genetics
information for melanoma research .
P04.140
A semi-automated unbiased differential methylation screening
assay for applications in cancer epigenetic research
V. V. Shkarupo 1,2 , A. S. Tanas 3 , E. B. Kuznetsova 1,4 , L. E. Zavalishina 5 , G. A.
Frank 5 , D. V. Zaletaev 1,4 , V. V. Strelnikov 1,4 ;
1 Research Centre for Medical Genetics, Moscow, Russian Federation, 2 Russian
State Medical University, Moscow, Russian Federation, 3 Sitronics Telecom
Solutions, Moscow, Russian Federation, 4 I.M.Sechenov Moscow Medical Academy,
Moscow, Russian Federation, 5 Moscow P.A.Gertsen Oncology Research
Institute, Moscow, Russian Federation.
Screening for differential methylation in tumor and control samples
is one of the powerful approaches towards the identification of novel
cancer related genes . Still there is no universal technique to screen
for differential methylation, although the spectrum of methylation detection
methods is vast . We have developed a synthetic highly effective,
unambiguous, unbiased, semi-automated screening technique to
serve as a possible basis for cancer epigenome research . Our differential
methylation screening approach is based on a fluorescent
methylation sensitive PCR with primers complementary to the adaptors
ligated to the DNA digestion fragments. These AIMS (amplification
of intermethylated sites) products are analyzed on automatic genetic
analyzers with single-nucleotide resolution using GeneScan software .
Specific software for automatic identification of differentially methylated
fragments have been developed as well as the software capable
of identifying the AIMS fragments in silico by comparison with available
genome databases . Additional computer service is available to
assist in experimentation design respective to specific applications.
With technical variations this approach may be elaborated to identify
novel targets of DNA methylation/demethylation as well as to roughly
characterize epigenomic status as a whole . We suggest that the assay
is universal and can be applied for screening differential methylation
in any disease where epigenetic component may be suspected . The
study is supported by Friends for an Earlier Breast Cancer Test Foundation,
USA.
P04.141
the investigation of tumor suppressor gene (P53) LOcUs-17
P13 in late stage Endometriosis patients by fluorescent in Situ
Hybridisation (Fish) technique
F. A. Teksen 1 , &. Keleş 2 , H. Şatıroğlu 3 ;
1 Faculty of Health Sciences, Ankara, Turkey, 2 Düzen Laboratory, Ankara, Turkey,
3 Ankara University Faculty Medicine, Obstetrics and Gynecology Department
and Biotechnology Institute, Ankara, Turkey.
In the study, the incidence of genetic alterations in p53 gene locus in
late stage endometriosis cases were investigated in comparison with
the endometrium and peripheral blood samples of the same patients
and the same tissues of normal healty women .
The study group was composed of six endometriosis samples (n:6) .
Control group included; normal endometrium samples (n:6, control I)
and peripheral blood lymphocytes (n:6, control II) of the same patients
with normal endometrium samples (n:5, control III) and peripheral
blood lymphocytes (n:5, control IV) of the healty women .
Frequency of monosomic, disomic and trisomic cells in controls and
endometriotic tissue specimens were determined by using Multicolor
Fluorescence in situ hybridization (FISH) technique and statisitically
analysed by Khi squared test .
In endometriosis samples the frequency of monosomy and trisomy
were 16 .4 % and 2 .1 % respectively for chromosome 17 p13 (p53)
locus . In endometrium samples (n:6) the frequency of trisomy was 3 .2
% . In peripheral blood lymphocytes (n:6) the frequency of trisomy was
0 .3 % for the same locus .
Finally, chromosome 17p13 (p53) locus genetic alterations were found
to be significantly greater (p