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, 2014 | OLOMOUC | THE CZECH REPUBLIC
ABS TRAC T BOOK
the near future, we will be facing several issues. One of them is
the problem of sufficient amount of material (e.g. in lung cancer
patients, where majority of cases is detected solely from small
endobronchial/transthoracic biopsies). Another is economical
issue, as the costs of such complex testing is steeply growing,
although it still represents much lower figures than the costs of
subsequent treatment based on the results of these tests. Still
another issue might be the sufficient number of pathologists
as the increasing precision and detailed growing number of
tests presumes much more efforts of pathologist resulting in
progressively increasing workload.
We live in an exciting period of history, when the new scien-
tific achievements do change the practice of everyday modern
medicine. However, if we will not identify and successfully solve
the potential issues, the future progress may be limited by
several factors. Modern pathology is a discipline which proved
to be an integral part of the current multidisciplinary approach
in diagnostics and treatment of the patients.
Supported by Charles University project PRVOUK P37/11
Molecular pathology of breast cancer progression
and the tumor microenvironment
Burkadze G.
Department of Pathology and Cytopathology,
Tbilisi State Medical University, Tbilisi, Georgia
Tumor progression – tumor metastatic and malignant phe-
notype – depends on interactions between tumor cells and
the tumor microenvironment. Different components of the
tumor microenvironment may have stimulatory or inhibitory
effects on tumor progression by regulating gene expression
profiles in tumor and stromal cells.
The importance of the microenvironment in breast carcino-
genesis is well-documented. Various experimental studies, as
well as observations on patients showed that stromal compo-
nent plays a crucial role during tumor development, and a wide
range of pathways have been implicated in cancer including
self-sufficiency in growth signals, insensitivity to antigrowth
signals, limitless replicative potential, evasion of apoptosis,
sustained angiogenesis, tissue invasion and metastasis. The
dynamic and reciprocal interactions between tumor cells
and cells of the microenvironment contribute critical events
to tumor evolution toward metastasis, and many cellular and
molecular elements of the microenvironment are emerging
as attractive targets for therapeutic strategies.
Agilent SureFISH – probes for rapid
identification of a wide range of chromosomal
aberrations across the genome
Halbhuber Z.
HPST s.r.o., Praha
Fluorescence
in situ
Hybridization (FISH) is a powerful tech-
nique for studying the structure, organization, and localization
of nucleic acids within individual cells. However, the use of FISH
has often been dependent upon access to cloned template
DNA for the generation of probes. Clones for specific regions
may be unavailable, or the genomic region of interest may
contain repetitive and other non-informative sequences which
can be problematic for FISH analysis. Agilent has leveraged
its ability to chemically synthesize DNA in massively parallel
reactions to produce libraries of oligonucleotides up to 200
bases in length that can be used to generate FISH probes.
The sequences of the oligonucleotides in these libraries are
selected
in silico
using empirically determined criteria so as
to avoid repetitive elements or regions homologous to other
non-targeted loci. Using oligonucleotide library-derived FISH
probes on DNA, human genomic regions as small as 1.8 kb
and as large as whole chromosomes can be visualized in both
metaphase and interphase cells using the same simple assay
protocol. Because of the inherent flexibility in our probe design
methods, we readily visualized regions rich in repeats and/or
GC content. Using oligonucleotide library-derived FISH probes
on RNA, Agilent has been able to detect the localization of
a variety of both coding and non-coding RNAs in fixed cells,
using both conventional fluorescence and structured illumi-
nation microscopy. Simultaneous hybridization of FISH probes
labeled with different fluorophores enables visualization of
multiple sequences at once. Using probes designed specifically
to transcribed vs. non-transcribed regions, Agilent has been
able to simultaneously detect DNA and RNA from the same
locus in the same FISH assay. Agilent’s oligo FISH methods are
readily compatible with the co-detection of cellular proteins
by immunocytochemistry. The ability to generate high perfor-
mance FISH probes using chemically synthesized oligo libraries
that can work flexibly with co-detection of other molecules
yields a valuable tool for studies of how localization of specific
nucleic acids impacts biological function.
