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

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28. S-ADENOSYLHOMOCYSTEINE HYDROLASE REGULATES C2-CERAMIDE INDUCED CELL DEATH VIA INHIBITION OF THE INDUCTION OF THE INTRINSIC APOPTOTIC PATHWAY Roman Hudec 1,3,* , Kozo Hamada 1,2 , Hideaki Ando 1,2 and Katsuhiko Mikoshiba 1,2 1 Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan 2 Calcium Oscillation Project, ICORP-SORST, Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan 3 Institute of Biochemistry, Nutrition and Health Protection, Department of Biochemistry and Microbiology, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinskeho 9, 812 37 Bratislava, Slovak Republic *roman.hudec@stuba.sk S-adenosylhomocysteine hydrolase (SAHH) is the rate limited enzyme of the global S-adenosylmethionin (SAM) dependent transmethylation of various biological substrates. Here we report, that SAHH and in particular the SAHH enzymatic activity, is essential for the C2-ceramide induced apoptosis. SAHH knock down by the specific siRNA, disabled the C2-ceramide induced methylation of the catalytic subunit of protein phosphatase 2A (PP2Ac) and therefore its activation. Such negative regulation led to inhibition of the C2- ceramide induced apoptosis via mitochondrial intrinsic pathway. The opposite situation has been observed by the transient transfection with the plasmid carried SAHH coding sequence. Introduced SAHH protein caused augmentation of the endogenous SAHH activity and subsequently caused elevation of the C2-ceramide induced apoptosis level. SAHH protein levels are in general low in the neoplastic tissues; establishment or reestablishment of its physiological level might be beneficial for the chemotherapy successfulness, since many of currently used chemotherapeutics act also via mechanisms involving the elevation of the endogenous ceramide levels and apoptosis induction. Acknowledgements This work was supported by grants from the Japan Society for the Promotion of Science (JSPS), the Ministry of Education, Science, Sports and Culture of Japan to K.M (20220007), and ICORP-SORST of JST. R.H. was a JSPS postdoctoral fellow. Analytical Cytometry VII 53
29. INFLUENCE OF THE LEVEL OF CD133 EXPRESSION ON ADHESION AND PROLIFERATION OF CANCER STEM-LIKE CELLS Radek Fedr 1# , Zuzana Pernicová 1,2# , Šárka Šimečková 1,3 , Veronika Toporcerová 1,3 , Alois Kozubík 1,3 , Karel Souček 1,2 * 1 Department of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic; 2 Center of Biomolecular and Cellular Engineering, International Clinical Research Center, St. Anne´s University Hospital Brno, Brno, Czech Republic; 3 Department of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic # equal contribution, *ksoucek@ibp.cz CD133 is a transmembrane glycoprotein that has been described as one of the typical cancer stem cells (CSC) marker in many types of tissue. However, the fact if the expression of CD133 unambiguously defines subpopulation of CSC is still controversial. Interestingly the function of CD133 is poorly described. Clonogenic assay is widely used in vitro for elucidating the proliferative capacity of every single cell in the population. Here, we used this assay to determine, if expression of cancer stem cell markers CD133 and CD44 correlates also with functional properties of cancer cells derived from conditional PTEN-deletion mouse model from androgen-independent tumor. Moreover, label-free real time cell analyzer xCELLigence was used to compare adhesion and proliferation of subpopulations with different expression of cancer stem cell markers. We found that CD44+/CD133high subpopulation of prostate cancer cells has significantly higher clonogenic capacity in comparison to CD44+/CD133low subpopulation. Moreover increased clonogenic capacity correlates with both increased adhesion and proliferation rate in CD133high subpopulation, as confirmed using label-free measurement with real time cell analyzer xCELLigence. Interestingly, fibronectin coating significantly increases adhesion of CD44+/CD133low and more significantly of CD44+/CD133high subpopulation. Blocking fibronectin motif by pre-treatment with synthetic RGDS peptide decreases ability of CD133high cells to adhere to fibronectin. We conclude that CD133 expression might contribute to the regulation of cell cycle and adhesion of prostate cancer cells. However further studies are necessary to describe mechanisms of these features. Acknowledgements This work was supported by grants IGA MZD NT13573-4/2012, AV ČR M200041203, OrganoNET (CZ.1.07/2.4.00/31.0245), HistoPARK (CZ.1.07/2.3.00/20.0185) and by project FNUSA-ICRC (no. CZ.1.05/1.1.00/02.0123) from the European Regional Development Fund. Institutional support was provided by the Academy of Sciences of the Czech Republic. 54 Analytical Cytometry VII
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stimulated cells. The physiological
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chemotherapeutic drug LA-12. These
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aberrant expression, localization a
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P21. ASSESSMENT OF MITOCHONDRIAL FU
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P23. PTEROSTILBENE: COMPARISON OF A
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Fam123b. Amer1 has been shown very
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In summary, we described different
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this process. Combined treatment wi
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Homolog 1) gene; previously identif
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was harmful neither alone nor in co
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P34. PHARMACOLOGICAL INHIBITION OF
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novel chemotherapeutics with specif
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P38. EVIDENCE OF ABNORMAL PERIPHERA
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acid to 5-lipoxygenase. Similarly t
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P41. WHY CAN WE SEE DIFFERENT RADIO
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P42. ELUCIDATION OF CROSSTALK BETWE
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P49. NEW ANALOGS OF RHODAMINE Jiři
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P51. FLUORESCENCE-LABELED FERROMAGN
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could be achieved via the activatio
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28. Pp. 1565-1570. 2008. Dearden C:
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cells (SFC) using IFN-γ Elispot in
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FACSCantoII flow cytometer (Becton
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P59. EARLY DIAGNOSTICS OF CARTILAGE
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cross-reacting group (CREG) and sha
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the frequency of autoimmune disease
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non-covalent bonds. For a solution
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P65. OVULATION STIMULATION PROTOCOL
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Acknowledgements This work was supp
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P68. SUBPOPULATIONS OF B LYMPHOCYTE
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triggers a process of normal blood
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3 Department of Internal Medicine -
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P73. THE V-ATPASE-INHIBITOR BAFILOM
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P75. 5-FLUOROURACIL-SELECTED TUMOUR
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We conclude that decrease in the po
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Acknowledgements This work was supp
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P79. NEW TYPE OF PHOTOSENSITIZER IS
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P83. DIFFERENTIATION OF NEURAL CRES
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given cell type are missing and the
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Our results show that both p38α +/
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P90. TOLL-LIKE RECEPTOR 2 TRACKS TH
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for chemokines in attracting “inf
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in immune organs (Bursa of Fabricii
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