ABSTRACTS – ORAL PRESENTATIONS - AMCA, spol. s r.o.

cell lines using multiple methodical approaches. Human metastatic cancer cell lines
were used as positive controls.
The goal of this work was to find an optimal functional assay which could be used for
subsequent functional assessment of specific molecules in cell migration and invasion.
To study cell migration, we performed chemotactic migration assays using transwell
chambers or real-time analysis with xCELLigence system. Alternatively, a “wound healing
- scratch assay” or agarose spot invasion assay was tested. Cell invasion was studied in a
similar setup to cell migration, after coating the membranes with ECM proteins such as
collagen or Matrigel. Activity of ECM degrading enzymes was analyzed by zymography
or a fluorescence microscopy-based gelatinase assay, expression of MMP proteins was
evaluated by western blotting.
Cell migration towards chemoattractant was enhanced by starving the cells in growth
factor depleted media. We observed a specific and robust activation of TGF-beta
signaling pathway under these conditions. These results were correlated with expression
of a set of secreted proteins on a cytokine array. Altogether, we observed that cancer
transformation of human benign prostate hyperplasia-derived BPH-1 cells is associated
with increased migration potential, but not with significant enhancement of cell invasion
associated with ECM protein degradation.
This work was supported by grant no. P301/12/P407 of the Czech Science Foundation;
by grant no. NT13573-4/2012 of the Ministry of Health of the Czech Republic; by the
Academy of Sciences of the Czech Republic, grant no. AV0Z50040702, and by the project
FNUSA-ICRC no. CZ.1.05/1.1.00/02.0123 from the European Regional Development
Fund.
References:
Slabakova, E., et al., TGF-beta1-induced EMT of non-transformed prostate hyperplasia
cells is characterized by early induction of SNAI2/Slug. Prostate, 2011.
P29. NKD1- CREER T2 MOUSE STRAIN: A NEW TOOL FOR SITE-SPECIFIC
RECOMBINATION IN WNT RESPONSIVE CELLS OF MOUSE INTESTINE AND LIVER
Fafílek Bohumil 1 , Stančíková Jitka 1,2 , Hlavatá Adéla 1,2 , Kořínek Vladimír 1
1
Laboratory of Cell and Developmental Biology, IMG AV CR (jitka.stancikova@img.cas.cz)
2
Faculty of Science, Charles University in Prague
Wnt signaling pathway plays a crucial role in ontogenesis and development of
all metazoans. In adult mammals, the Wnt signaling pathway is required for the
maintenance of the intestinal homeostasis and establishment of proper hepatic zonation.
In contrary to that, aberrant activation of the Wnt pathway leads to neoplasia and cancer
development, notably in the intestine and liver.
To investigate the role of the Wnt pathway in gut epithelium homeostasis and its
malignant transformation we employed chromatin immunoprecipitation method (ChIP)
in combination with DNA microarrays (so-called ChIP-on-chip) to identify genes regulated
by the Wnt signaling. One of the most prominent targets was the NKD1 (Naked Cuticle
Analytical Cytometry VII 121