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

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cells (LSC) are present in CD45+34+38-26+ subpopulation only. Loss of expression of CD26 on Ph - cells from patients with pattern 2 appears to be specific for after-therapy phase of the disease. On the other hand, not all CD45 + 34 + 38 - 26 + cells were found Ph positive. Our findings confirm that CD26 antigen is useful for discrimination between Ph + and Ph - cells in exact disease phase and/or pattern-dependent only. However, considering easy identification of chronic myeloid stem cells by Ph positivity in CD45 + 34 + 38 - 26 - or CD45 + 34 + 38 - 26 + populations, it is obvious that precise separation LSC from HSC in target populations using CD26 is not possible in all settings. Despite these facts, this approach opens up a new possibilities for CML study. Acknowledgements Supported by MH CZ - DRO (FNBr, 65269705) References Gerber JM, Gucwa JL, Esopi D, Gurel M, Haffner MC, Vala M, Nelson WG, Jones RJ, Yegnasubramanian S.: Genome-wide comparison of the transcriptomes of highly enriched normal and chronic myeloid leukemia stem and progenitor cell populations. Oncotarget. 2013 Janssen JJ, Deenik W, Smolders KG, van Kuijk BJ, Pouwels W, Kelder A, Cornelissen JJ, Schuurhuis GJ, Ossenkoppele GJ.: Residual normal stem cells can be detected in newly diagnosed chronic myeloid leukemia patients by a new flow cytometric approach and predict for optimal response to imatinib. Leukemia 2012 64. EPITHELIAL-TO-MESENCHYMAL TRANSITION IN SUBPOPULATION OF MOUSE PROSTATE STEM CELLS AND CANCER STEM-LIKE CELLS Zuzana Pernicová 1,2 , Eva Slabáková 1,2 , Radek Fedr 1 , Maximilian Marhold 3 , Erwin Tomasich 3 , Peter Horak 3 , Alois Kozubík 1,4 , Karel Souček 1,2 1 Department of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic; E-mail: pernicova@ibp.cz 2 Center of Biomolecular and Cellular Engineering, International Clinical Research Center, St. Anne‘s University Hospital Brno, Brno, Czech Republic 3 Department of Internal Medicine I – Oncology, Comprehensive Cancer Center, Medical University of Vienna, Wien, Austria 4 Department of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic Epithelial-to-mesenchymal transition (EMT) is an important process during embryogenesis, which may be re-activated in several pathophysiological processes including tumor progression. In several models, normal cells or cancer cells undergoing EMT display properties and characteristics of stem cells (Kong et al., 2011; Mani et al., 2008). In our work, we aimed to identify EMT transcription factors with altered expression in subpopulation of mouse prostate stem cells or cancer stem cells, respectively, to prove involvement of EMT in biology of stem cells also in mouse prostate. There are several Analytical Cytometry VII 81
approaches how to detect and isolate mouse prostate stem cells and cancer stem cells, respectively. According to published results (Goldstein et al., 2008), using multicolor flow cytometry we detected putative mouse prostate adult stem cells with phenotype Lin - /Sca1 + /CD49f + /Trop-2 + in normal prostate and prostate cancer. To elucidate whether subpopulation of cells expressing stem cells markers display changes in levels of EMT markers and regulators, using high speed sorter we isolated subpopulations of cells with phenotype Lin - /Sca1 + /CD49f + /Trop-2 + and Lin - /Sca-1 - /CD49f - . Further we introduced a PCR method for detection of mRNA level from limited number of sorted cells and compared mRNA levels of EMT regulators (SNAI1, SNAI2, TWIST2, ZEB1, ZEB2), and markers of differentiation (VIM and CDH1) in these subpopulations. Our results show that EMT transcription factor SNAI2/Slug is strongly up-regulated in subpopulation of both mouse prostate stem cells and cancer stem cells with phenotype Lin - /Sca1 + /CD49f + /Trop2 + . Interestingly, this was not accompanied with up-regulation of EMT markers, since vimentin was found to be strongly down-regulated in both mouse prostate stem cells and cancer stem cells subpopulations. Further we aimed to confirm this Slug up-regulation also on protein level using multicolor flow cytometry protocol for simultaneous detection of selected surface and intracellular markers. Taken together our results show that subpopulation of cells with stem cell characteristics can be detected in both wt prostate and prostate cancer using combination of surface markers Sca-1, CD49f and Trop-2. Moreover, we found out that transcription factor and regulator of EMT Slug is strongly up-regulated in this subpopulation of putative prostate stem cells / cancer stem cells. Acknowledgements This work was supported by grants IGA MZD NT13573-4/2012, AV ČR M200041203, GA ČR P301/12/P407, 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. References Goldstein, A. S., Lawson, D. A., Cheng, D., Sun, W., Garraway, I. P., and Witte, O. N.Trop2 identifies a subpopulation of murine and human prostate basal cells with stem cell characteristics: Proc Natl Acad Sci U S A, v. 105, p. 20882-7, 2008. Kong, D., Li, Y., Wang, Z., and Sarkar, F. H. Cancer Stem Cells and Epithelial-to-Mesenchymal Transition (EMT)-Phenotypic Cells: Are They Cousins or Twins?: Cancers (Basel), v. 3, p. 716-729, 2011. Mani, S. A., Guo, W., Liao, M. J., Eaton, E. N., Ayyanan, A., Zhou, A. Y., Brooks, M., Reinhard, F., Zhang, C. C., Shipitsin, M., Campbell, L. L., Polyak, K., Brisken, C., Yang, J., and Weinberg, R. A. The epithelial-mesenchymal transition generates cells with properties of stem cells: Cell, v. 133, p. 704-15, 2008. 82 Analytical Cytometry VII
Page 1 and 2: ABSTRACTS - ORAL PRESENTATIONS 14.
Page 3 and 4: and (iii) siRNA-mediated knockdown
Page 5 and 6: 25. PROFILING OF CHILDHOOD ACUTE LE
Page 7 and 8: more dominantly than Th1 and CD4 ce
Page 9 and 10: 29. INFLUENCE OF THE LEVEL OF CD133
Page 11 and 12: 36. B-CELL IMMUNOPHENOTYPING OF LAR
Page 13 and 14: possible mechanism behind risk alle
Page 15 and 16: isotype controls and FMO for CD14 a
Page 17 and 18: NEP developmental stages were disti
Page 19 and 20: 50. THE PROGRESSION OF HIV INFECTIO
Page 21 and 22: 52. PRŮKAZ REGULAČNÍCH T LYMFOCY
Page 23 and 24: values when analysing paediatric ly
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Page 27 and 28: for application of proton therapy,
Page 29 and 30: effects will be also characterized
Page 31 and 32: Although multiple signalling pathwa
Page 33 and 34: an ability to recreate the tumour f
Page 35: incubated with non-filtered superna
Page 39 and 40: progress in development of automate
Page 41 and 42: tissues is characteristic for a num
Page 43 and 44: kappaB. Beside that, we found OANO
Page 45 and 46: therapy in patients (Calderwood et
Page 47 and 48: difference in the effect caused eit
Page 49 and 50: 4 CIC bioGUNE Technology Park of Bi
Page 51 and 52: P8. PROADIFEN (SKF-525A) ATTENUATES
Page 53 and 54: models. As manifested by compromise
Page 55 and 56: P11. NEW BIOACTIVE AND FLUORESCENT
Page 57 and 58: Acetylation of histones, along with
Page 59 and 60: stimulated cells. The physiological
Page 61 and 62: h induced very minor manifestations
Page 63 and 64: chemotherapeutic drug LA-12. These
Page 65 and 66: aberrant expression, localization a
Page 67 and 68: P21. ASSESSMENT OF MITOCHONDRIAL FU
Page 69 and 70: P23. PTEROSTILBENE: COMPARISON OF A
Page 71 and 72: Fam123b. Amer1 has been shown very
Page 73 and 74: In summary, we described different
Page 75 and 76: this process. Combined treatment wi
Page 77 and 78: Homolog 1) gene; previously identif
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Page 81 and 82: was harmful neither alone nor in co
Page 83 and 84: P34. PHARMACOLOGICAL INHIBITION OF
Page 85 and 86: 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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P44. INSTRUMENT SETTINGS FOR EUROFL
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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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LD11015, from GAČR 13-29358S, and
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P83. DIFFERENTIATION OF NEURAL CRES
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given cell type are missing and the
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and HistoPARK (CZ.1.07/2.3.00/20.01
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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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ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.