53
APRIL 24
TH
– 26
TH
, 2014 | OLOMOUC | THE CZECH REPUBLIC
ABS TRAC T BOOK
tumor 1, gen klíčový při vývoji urogenitálního traktu a muto-
vaný u části pacientů s Wilmsovým tumorem) a PYCARD (PYD
and CARD domain containing, protein zprostředkující tvorbu
velkých komplexů signálních drah apoptózy a zánětu cestou
aktivace kaspáz) byla zjištěna výlučně ve vzorcích nádorové
tkáně. Důsledky těchto epigenetických změn pro patogenezi
HCC jsou předmětem dalšího zkoumání.
Podpořeno MZ ČR – RVO (FNHK, 00179906), MŠMT ČR
CZ.1.07/2.3.00/20.0019, SVV LF UK HK 266902 a 264902.
9) Implementation of MassArray® technology
for determination of SLCO1B1 gene variant
associated with adverse effect of statin therapy
Pašková L.
1,3
, Petřková J.
1,2,3
, Nosek Z.
1,3
, Petřek M.
1,3
1
Laboratory of Cardiogenomics, Department of Clinical
and Molecular Pathology
2
Ist Clinic of Internal Medicine Cardiology, Faculty
Hospital Olomouc
3
Immunogenomics, Institute of Molecular and
Translational Medicine, Faculty of Medicine and
Dentistry, Palacky University Olomouc
Introduction:
statins are highly effective drugs in lowering
plasmatic concentration of LDL-cholesterol. Aggressive treat-
ment with statins reduces the risk of cardiovascular events in
patients with coronary artery disease. Organic anion transport-
ing polypeptide 1B1 (OATP1B1) encoded by the gene
SLCO1B1
plays an important role in statin metabolism. The presence
of SNP rs4149056 (521T>C) in the
SLCO1B1
gene has been as-
sociated with increased risk of statin-induced adverse effects
such as myopathy. Therefore we aimed at implementation of
a reliable and effective genotyping method for rs4149056 de-
termination using Sequenom
®
(MassArray
®
) technology, which
combines multiplex PCR amplification with MALDI-TOF mass
spectrometry detection.
Method:
Specific primers for genotyping of SNP rs4149056
were designed by Assay Design Suite (Sequenom
®
). DNA
samples obtained from 123 patients with acute myocardi-
al infarction were processed using iPLEX
®
Gold Reagent Kit
(Sequenom
®
). The results of genotyping were obtained in
36-48 hours after blood sampling. The allele and genotype
frequencies of rs4149056 were determined by direct count-
ing and compared with those reported previously for other
European populations.
Results:
The frequency of the rs4149056 low activity C
allele was 17.5 %, which is similar to the frequency (18.0 %)
determined in European populations. The distribution of indi-
vidual genotypes was TT = 69.1 %, TC = 26.8 % and CC = 4.1 %,
which complied with Hardy-Weinberg equilibrium (P>0.05).
The results show that 31.0 % of our patients carried the“risk” C
allele in their genotype.
Conclusion:
The distribution of
SLCO1B1
(rs4149056) SNP
in the investigated Czech population does not differ from
the other European populations. The MassArray
®
technol-
ogy enables accurate and relatively fast determination of the
SLCO1B1
polymorphism. Thereby this methodology may be
recommended for early identification of patients in risk of ad-
verse reactions to statin treatment and for its individualization.
Grant support: IGA PU LF 2014_012 and CZ.1.05/2.1.00/01.0030
10) Somatic mutations in IDH1 and IDH2 genes
in gliomas
Urbanovská I.
1,2,3
, Konvalinka D.
1
, Šimová J.
1,2
,
Kubová B.
1
, Měch R.
1
, Uvírová M.
1,2,3
, Tomanová R.
2,4
,
Buzrla P.
4
, Paleček T.
5
, Drábek J.
3
, Dvořáčková J.
2,4
1
AGEL Research and Training Institute – Ostrava-
Vítkovice Branch, CGB Laboratory
2
Department of Biomedical Sciences Faculty of Medicine,
University of Ostrava, Ostrava
3
Institute of Molecular and Translational Medicine Faculty
of Medicine, Palacký University, Olomouc
4
Institute of Pathology, University Hospital in Ostrava
5
Neurosurgery Clinic, University Hospital in Ostrava
Primary brain tumors account for about 2 % of all adult ma-
lignancies. The incidence of primary brain tumors has nearby
doubled within the past 30 years and was 7.95 per 100 000
inhabitants in the Czech Republic (in 2010). The most common
brain tumors are gliomas and meningiomas. Gliomas include
tumors of various types and grades.
IDH1 and IDH2 genes are coding isocitrate dehydrogena-
ses, which are members of isocitrate dehydrogenase family,
catalysing the oxidative decarboxylation of isocitrate to α-ke-
toglutarate, resulting in the production of NADPH.
Mutations in IDH1 and IDH2 genes are frequent in low-gra-
de astrocytic and oligodendroglial tumors and in secondary
glioblastomas. IDH mutations are rare or absent in other types
of glial tumors, such as pilocytic astrocytomas and ependy-
momas. These mutations are associated with more favourable
prognosis and prolonged overall survival in glioblastoma pati-
ents. IDH1 mutations are located in codon 132 with the most
frequent R132H mutation, which was observed in 83–91 % of
all mutations.
Mutations in IDH2 gene are rare and occur almost exclusi-
vely in codon 172.
In our study mutation R132H in IDH1 gene was determined
by immunohistochemistry with IDH1 R132HMouse anti Human
unconj. antibody (Dianova GmbH) and further investigated
for presence of mutations in IDH1 and IDH2 genes in glioma
patiens by molecular genetic methods (SNaPshot assay, se-
quencing analysis).
Determination of IDH1/2 mutations has prognostic and
diagnostic value and can contribute to the exact differentiation
between histological types of gliomas.
