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

in MEF cells reveals requirement of the interaction between AMER1 and β-arrestin for
Wnt3a-induced AMER1 dynamics. (A-C) WT and β-arr1/2 DKO MEFs were transfected
with EGFP-AMER1 (A, B) or EGFP-AMER1(2-838) (C) and membrane dynamics of the
constructs was analyzed by Fluorescence Recovery After Photobleaching (FRAP) method.
Example of cells subjected to FRAP is given on the left, squares indicate bleached regions.
On the right statistical analysis of FRAP experiment using WT and β-arr1/2 DKO MEFs
stimulated with PBS or Wnt3a is shown. The graphs show mean values + s.e.m., and
the best fitting curve model, which was used for calculation of mobile pool of EGFP-
AMER1 (% of fluorescence recovered) and of the recovery halftime (T1/2). N – number
of analyzed cells.
P25. DETECTION OF CANCER STEM CELL MARKERS AND ANALYSIS OF EPITHELIAL-TO-
MESENCHYMAL TRANSITION IN PROSTATE BENIGN AND CANCER CELL LINES
Eva Sedlmaierová 1,2* , Zuzana Pernicová 1,3 , Šárka Šimečková 1,2 , Eva Slabáková 1,3 , Radek
Fedr 1 , Alois Kozubík 1,2 , Karel Souček 1,3*
1
Department of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech
Republic, Brno, Czech Republic;
2
Department of Experimental Biology, Faculty of Science, Masaryk University, Brno,
Czech Republic;
3
Center of Biomolecular and Cellular Engineering, International Clinical Research Center,
St. Anne´s University Hospital Brno, Brno, Czech Republic
*Correspondence to: sedlmaierova@ibp.cz, ksoucek@ibp.cz
Cancer stem cells (CSCs) have long been implicated in numerous tumour formations
and therapy resistance including prostate cancer disease. Importantly, mechanisms of
their origin have not been elucidated in prostate cancer yet. It has been demonstrated
that epithelial-to-mesenchymal transition (EMT) is phenomenon which can lead to the
acquisition of stem cell properties in various types of cancer. This was first discovered
by research on immortalized human mammary epithelial cells, where induction of EMT
resulted in the acquisition of mesenchymal properties and in the expression of stem
cell markers. Furthermore those cells had properties similar to mammary epithelial
stem cells (Mani et al., 2008). In prostate cancer, cells with EMT phenotype have been
shown to express stemness factors and to have increased clonogenic capacity (Kong et
al., 2010). However phenotypic heterogeneity and EMT status of routinely cultivated
prostate cell lines remains unknown.
In this study we aimed to screen surface expression of selected CSCs markers (Trop2,
CD133, CD44, CD24 and CD49f) in a panel of prostate benign and cancer cell lines using
multicolour flow cytometry. We found out that each cell line express different set and
combinations of these CSCs markers. Next, to elucidate the EMT status in putative
CSCs subpopulations, we combined analysis of selected surface stem cell markers with
detection of regulators and markers of EMT (Snail, Slug, E-cadherin, N-cadherin) in
CD24/CD44 subpopulation of PC-3 and DU-145 cells.
Analytical Cytometry VII 117