2009 Vienna - European Society of Human Genetics

Cancer genetics
from patients with benign prostatic hyperplasia, but it was found in 25
( 86,2%) biological samples from patients with prostate adenocarcinoma.
Conclusions: Analysis by methylation specific PCR of GSTP1 promoter
hypermethylation provides a specific tool for the early molecular
diagnosis of prostate cancer in blood and tissue samples.
P06.076
Y chromosome haplogroup R1a is associated with prostate
cancer risk among macedonian males
D. Plaseska-Karanfilska 1 , P. Noveski 1 , N. Matevska 2 , A. Dimovski 2 , G. D.
Efremov 1 ;
1 Macedonian Academy of Sciences and Arts, Research Centre for Genetic
Engineering and Biotechnology, Skopje, Macedonia, The Former Yugoslav
Republic of, 2 Faculty of Farmacy, Center for Biomolecular Sciences, Skopje,
Macedonia, The Former Yugoslav Republic of.
Prostate cancer (PC) is one of the most common male-specific cancers.
Its incidence varies considerably between populations. Recent
surveys suggest that PC is influenced by both genetic and environmental
factors, although the etiology of the disease remains unknown
in the majority of cases. Certain Y chromosomal lineages have been
suggested to predispose individuals to prostate cancer in Japanese
population, but no association has been found among Korean and
Swedish patients. The aim of this study was to investigate the association
between Y chromosomal haplogroups and predisposition to
prostate cancer in Macedonian men. We studied 84 PC patients and
126 males from the general population of Macedonian ethnic origin. A
total of 28 markers have been studied by multiplex PCR and SNaPshot
analysis. Nineteen different Y haplogroups were determined; the
most frequent being I1b-P37b, E3b1-M78, R1a-SRY 1532, R1b-P25
and J2b1a-M241. The frequency of R1a was significantly higher in
PC patients (20.2%) in comparison with the controls (9.5%) [p=0.027;
OR=2.41 (1.09-5.36)]. When stratified according to age, even stronger
association was observed between haplogroup R1a and prostate
cancer in patients of >65 years of age [p=0.004; OR=3.24 (1.41-7.46)].
Our results suggest that Y chromosome haplogroup R1a is associated
with an increased prostate cancer risk in Macedonian men.
P06.077
multifaceted preventive effects of single agent quercetin on a
human prostate adenocarcinoma cell line (Pc-3): implications to
nutritional transcriptomics and multi-target therapy
M. Momeny1 , N. Motamed2 , N. Kazemialiakbar2 , M. Yaseri1 , M. Yousefi1 , S.
Hashemi1 , M. R. Noori-Daloii1 ;
1 2 Tehran Univ. of Medical Sciences, Tehran, Islamic Republic of Iran, University
of Tehran, Tehran, Islamic Republic of Iran.
The aim of the present study is to evaluate the effects of quercetin, a
dietary flavonoid, on human prostate adenocarcinoma PC-3 cells. Lactate
dehydrogenase (LDH) release, microculture tetrazolium test (MTT
assay) and real-time PCR array were employed to assess the influences
of quercetin on cell cytotoxicity, cell proliferation and expression
of various genes in PC-3 cell line. Quercetin inhibited cell growth and
proliferation and modulated the expression of genes involved in DNA
repair, matrix degradation and tumor invasion, angiogenesis, apoptosis,
cell cycle, metabolism and glycolysis. More importantly, quercetin
inhibited the expression of genes responsible for progression from the
androgen deprivation-responsive stage to the hormone deprivation refractory
phase. In addition, no cytotoxicity of quercetin on PC-3 cells
was observed. Taken together, as shown by the issues of the current
study for the first time, the manifold inhibitory impacts of quercetin on
PC-3 cells may introduce quercetin as an efficacious “magic shotgun”
in order to be used in the future nutritional transcriptomic investigations
and multi-target therapy to overcome the therapeutic impediments in
crusade against prostate cancer.
P06.078
Low prevalence of PtEN mutations in a sample of italian
patients with cowden or cowden-like syndrome
L. M. Pradella 1 , C. Rossi 1 , A. Selicorni 2 , L. F. Pennisi 1 , G. Romeo 1 , D. Turchetti
1 ;
1 Cattedra e UO di Genetica Medica, Università di Bologna- Policlinico S.Orsola-
Malpighi, Bologna, Italy, 2 Ambulatorio di Genetica Clinica,Clinica Pediatrica
Università di Milano, Milano, Italy.
Cowden Syndrome (CS) is characterized by multiple hamartomatous
and neoplastic lesions. 80-85% of CS patients are reported to carry
detectable mutations in the PTEN gene. Recently, mutations in SDHB
and SDHD genes have been described in some PTEN-negative CS
patients. PTEN mutations have been also detected in about 60% of
patients with Bannayan-Riley-Ruvalcaba Syndrome (BRRS), whose
phenotype is partially overlapping that of CS.
We describe our preliminary experience of genetic testing in patients
with features of CS or BRRS. Overall, we tested 15 patients: 4 fulfilled
the criteria for the diagnosis of CS, 3 were affected by BRRS, whereas
the remaining 8 had some criteria for CS and were therefore classified
as CS-like.
Mutational analysis of PTEN was performed with automated direct
sequencing using a multistep approach: first, exons 2 to 7 (containing
80% of the described mutations) and their flanking regions were
analized. If no mutations were detected, the analysis was extended to
exons 1, 8 and 9 and to the PTEN promoter region. PTEN-negative CS
and CS-like patients were screened for mutations in the 4 coding exons
of SDHD and the 8 coding exons of SDHB and flanking regions.
Only in one CS patients a PTEN mutation was detected, which had
not been reported before; no PTEN, SDHB or SDHD mutations were
detected in the other CS, in CS-like and in BBRS patients. Based on
these preliminary results, the genetic bases of CS and BBRS in our
population appear to differ from those described in other populations.
P06.079
qPcR-HRm : a new approach in the screening of both point
mutation and large rearrangement - application to oncogenetic
C. Lefol1 , E. Rouleau1 , L. Demange1 , C. Nogues1 , V. Bourdon2 , F. Coulet3 , F.
Soubrier3 , H. Sobol2 , I. Bieche1 , S. Olschwang2 , R. Lidereau1 ;
1 2 Centre René Huguenin, St Cloud, France, Institut Paoli Calmettes, Marseille,
France, 3Hôpital Pitié Salpêtrière, Paris, France.
In germline oncogenetic diseases, deleterious mutations, point mutations
and large rearrangements are responsible of the inactivation in
tumor suppressor genes as BRCA1/BRCA2 in breast cancer or MLH1/
MSH2 in colorectal cancer. Until now, two different techniques were
required to fully pre-screen those genes. We propose a new approach
qPCR-HRM that combines quantitative PCR (qPCR) and high resolution
melting curve analysis (HRM). This approach change also the
way to perform high melting curve analysis. We illustrate this with our
experience in the MLH1 gene. 76 patients were fully pre-scanned for
mutation in MLH1 including 14 wild-type patients and 62 patients with
known mutations (57 point mutations and 5 rearrangements). Moreover,
a blind screening of 55 samples in triplicate was performed to
assess sensitivity and sensibility in comparison to dHPLC-MLPA.
In the 131 patients, all the mutations detected by dHPLC+MLPA, either
point mutations or rearrangements, were detected successfully
with qPCR-HRM. The sensitivity was similar to dHPLC. However, the
replicates in qPCR-HRM improve drastically the specificity. In the blind
screening, without considering the triplicate, there were 170 false positives
against 6 false positives with the triplicate (from a unique sample
with obvious DNA quality problems).
With qPCR-HRM, pre-screening for point mutations and large rearrangements
are realised in one tube and one step in a single machine
without use of an automatic sequencer in the pre-screening process.
The replicate approach increase the specifity of the HRM curve analysis.
qPCR-HRM outperformed other techniques in term of rapidity and
amount of data provided.
P06.080
Germline mutation in RAP80 impairs DNA damage response
function
J. Nikkilä 1 , K. A. Coleman 2 , D. Morrissey 2 , K. Pylkäs 1 , H. Erkko 1 , T. E. Messick
2 , S. Karppinen 1 , A. Amelina 1 , R. Winqvist 1 , R. A. Greenberg 2,3 ;
1 Laboratory of Cancer Genetics, Dept of clinical genetics and Biocenter Oulu,
University of Oulu, Oulu university hospital, Oulu, Finland, 2 Department of
Cancer Biology, Abramson Family Cancer Research Institute, University of
Pennsylvania School of Medicine., Pennsylvania, PA, United States, 3 Department
of Pathology, Abramson Family Cancer Research Institute, University of
Pennsylvania School of Medicine., Pennsylvania, PA, United States.
Background: 5-10% of all breast cancers stem from hereditary predisposition
to the disease. Mutations in two major susceptibility genes