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

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Procházka P, Libra A, Zemanová Z, Hřebačková J, Poljaková J, Hraběta J, Bunček M, Stiborová M, Eckschlager T. Mechanisms of ellipticine-mediated resistance in UKF-NB-4 neuroblastoma cells. Cancer Sci. 2012;103:334-41. Spugnini EP, Citro G, Fais S. Proton pump inhibitors as anti vacuolar-ATPases drugs: a novel anticancer strategy. J Exp Clin Cancer Res. 2010;29:44. P74. NESTIN EXPRESSION IN LEUKEMIA CELLS. Tomáš Loja 1,2 , Marek Borský 1 , Tomáš Bernard 1 , Michael Doubek 1,2 1 Department of Internal Medicine - Hematology and Oncology, University Hospital Brno and Faculty of Medicine, Masaryk University, Czech Republic; tomas.loja@gmail.com 2 CEITEC - Central European Institute of Technology, Masaryk University, Czech Republic Protein nestin is considered a marker of immature (stem/progenitor) cells and its expression occurs during mammalian embryogenesis. Re-expression of nestin in adults occurs in various pathologic conditions; neoplastic transformatio is one of them. Nestin has been detected in whole range of solid tumors where its expression correlates with tumor malignancy and invasiveness - nestin expression levels are higher in tumors with high grades than in those with low grades. Therefore, nestin is thought to be negative prognostic marker. In addition, some nestin-positive populations of tumor cells have the same features as cancer stem cells (CSCs) responsible for tumor relapse according to CSCs hypothesis. So far it is very little known about nestin expression in leukemia cells. Our pilot data proved nestin in tumor cells of various leukemia and lymphoma cell lines, as well as in patients with chronic lymphocytic leukemia (CLL) and other hematooncological diagnoses. CLL, the most common adult leukemia in Western countries, is a progressive hematopoietic disorder characterized by clonal proliferation and accumulation of neoplastic mature-appearing B-lymphocytes in the blood, bone marrow, spleen, and lymph nodes. Malignant lymphoproliferative diseases represent a group of cancers with high incidence, variable biological behavior and prognosis, therefore, attention is paid to new prognostic factors, which would allow to stratify patients according to their individual risk factors and to optimize patients treatment. Nestin expression level is linked to tumor subtypes which are less differentiated, more aggressive, and have poorer prognosis; therefore, determination of nestin expression in CLL patients as well as in other hematological malignancies could be used to estimate the biological behavior. In addition, combined analysis of nestin and other diagnostic/ prognostic factors may help to improve stratification and therapy of oncologic patients. Acknowledgements This work was supported by the project (Ministry of Health, Czech Republic) for conceptual development of research organization 65269705 (University Hospital Brno, Brno, Czech Republic). Analytical Cytometry VII 177
P75. 5-FLUOROURACIL-SELECTED TUMOUR CELLS EXERT CROSS-RESISTANCE WHICH CAN BE MODULATED BY PHARMACOLOGICAL AND MOLECULAR INHIBITION OF ALDEHYDE DEHYDROGENASE Miroslava Matuskova, Erika Durinikova, Lucia Kucerova, Zuzana Kozovska Cancer Research Institute of Slovak Academy of Sciences, Bratislava Slovak Republic One of the obstacles of effective cancer treatment is selection for chemo-resistant subpopulations of malignant cell during chemotherapy. Innovative approaches need to be developed to affect malignant cells resistant to conventional treatment. Chemo-resistant tumour cells use more mechanisms to escape cytotoxic effect of drugs. Increased expression of aldehyde dehydrogenase (ALDH), particularly 1A1 isoform (ALDH1A1) is also involved in drug resistance (Januchowski et al., 2013). We aimed to evaluate the sensitivity of 5-Fluorouracil (5-FU) resistant tumour cells to panel of drugs, and prove the effect of inhibition of ALDH to evaluate its role in response to chemotherapy. Model chemo-resistant tumour cell line HT-29/EGFP/FUR proliferating in clinically relevant plasma concentration of 5-FU was derived from human colon carcinoma cells HT-29 by long-term cultivation in sequentially increasing 5-FU concentration. Tumour cells were retrovirally transduced with enhanced green fluorescent protein (EGFP) for unequivocal identification in cytometric experiments and evaluation of sensitivity to chemotherapy treatment by fluorimetric assay. Immunophenotype, ALDH activity and apoptosis were determined by flow cytometry. Kinetic experiments were performed using Incucyte device. Gene expression was evaluated by qPCR, siRNA was delivered by nucleofection. Athymic mice were used for experiments in vivo. We demonstrated altered expression of enzymes involved in nucleotide metabolism and transporter protein ABCC4. Changes correlated with their suggested role in 5-FU resistance. Differences in expression of stem cells` markers ALDH1A1 and CD133 were observed. We expected increased expression of ALDH1A1 enzyme in chemo-resistant derivate of HT-29 cells. The functional assay ALDEFLUOR confirmed increased activity of ALDH however ALDH1A1 – specific qPCR showed decreased expression of 1A1 isoform. Lower expression of ALDH1 was also confirmed on protein level by Western-blotting. Other ALDH isoforms have probably increased expression. HT-29/EGFP/FUR cells were cross-resistant to Cis-Platin, Paclitaxel, Doxorubicine and Cyclofosamide, .which are structurally and functionally unrelated to 5-FU. HT-29/EGFP/FUR derived xenografts maintained their chemo-resistance for several months without selection pressure of 5-FU, but exerted lower tumorigenicity. MSC co-injection had tumour-favouring potential on the HT-29/EGFP/FUR xenograft growth. Immunomagnetically separated CD133-positive subpopulation showed decreased sensitivity to chemotherapy. ALDH activity was pharmacologically inhibited by diethylaminobenzaldehyde (DEAB) or by RNA interference using set of ALDH1A1 specific small interfering RNA (siRNA). Nucloefection of ALDH1A1 siRNA has short term effect on target genes’ expression. Decreased level of mRNA was observed up to 72 hours post transfection. Tumour cells with inhibited ALDH activity exerted altered sensitivity to 5-FU, Cis-platin and oxidative stress. 178 Analytical Cytometry VII
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ABSTRACTS - ORAL PRESENTATIONS 14.
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and (iii) siRNA-mediated knockdown
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25. PROFILING OF CHILDHOOD ACUTE LE
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more dominantly than Th1 and CD4 ce
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29. INFLUENCE OF THE LEVEL OF CD133
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36. B-CELL IMMUNOPHENOTYPING OF LAR
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possible mechanism behind risk alle
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isotype controls and FMO for CD14 a
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NEP developmental stages were disti
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50. THE PROGRESSION OF HIV INFECTIO
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52. PRŮKAZ REGULAČNÍCH T LYMFOCY
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values when analysing paediatric ly
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42,0) vs. 1,5 (0,0-28), p
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for application of proton therapy,
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effects will be also characterized
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Although multiple signalling pathwa
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an ability to recreate the tumour f
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incubated with non-filtered superna
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approaches how to detect and isolat
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progress in development of automate
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tissues is characteristic for a num
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kappaB. Beside that, we found OANO
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therapy in patients (Calderwood et
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difference in the effect caused eit
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4 CIC bioGUNE Technology Park of Bi
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P8. PROADIFEN (SKF-525A) ATTENUATES
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models. As manifested by compromise
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P11. NEW BIOACTIVE AND FLUORESCENT
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Acetylation of histones, along with
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stimulated cells. The physiological
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h induced very minor manifestations
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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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egulate expression of different gen
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Page 83 and 84: P34. PHARMACOLOGICAL INHIBITION OF
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Page 87 and 88: P38. EVIDENCE OF ABNORMAL PERIPHERA
Page 89 and 90: acid to 5-lipoxygenase. Similarly t
Page 91 and 92: P41. WHY CAN WE SEE DIFFERENT RADIO
Page 93 and 94: P42. ELUCIDATION OF CROSSTALK BETWE
Page 95 and 96: P44. INSTRUMENT SETTINGS FOR EUROFL
Page 97 and 98: e used to cultivate endothelial cel
Page 99 and 100: concentrations 0.15, 0.2, 0.3 and 0
Page 101 and 102: P49. NEW ANALOGS OF RHODAMINE Jiři
Page 103 and 104: P51. FLUORESCENCE-LABELED FERROMAGN
Page 105 and 106: could be achieved via the activatio
Page 107 and 108: 28. Pp. 1565-1570. 2008. Dearden C:
Page 109 and 110: cells (SFC) using IFN-γ Elispot in
Page 111 and 112: FACSCantoII flow cytometer (Becton
Page 113 and 114: P59. EARLY DIAGNOSTICS OF CARTILAGE
Page 115 and 116: cross-reacting group (CREG) and sha
Page 117 and 118: the frequency of autoimmune disease
Page 119 and 120: non-covalent bonds. For a solution
Page 121 and 122: P65. OVULATION STIMULATION PROTOCOL
Page 123 and 124: Acknowledgements This work was supp
Page 125 and 126: P68. SUBPOPULATIONS OF B LYMPHOCYTE
Page 127 and 128: triggers a process of normal blood
Page 129 and 130: 3 Department of Internal Medicine -
Page 131: P73. THE V-ATPASE-INHIBITOR BAFILOM
Page 135 and 136: We conclude that decrease in the po
Page 137 and 138: Acknowledgements This work was supp
Page 139 and 140: P79. NEW TYPE OF PHOTOSENSITIZER IS
Page 141 and 142: LD11015, from GAČR 13-29358S, and
Page 143 and 144: P83. DIFFERENTIATION OF NEURAL CRES
Page 145 and 146: given cell type are missing and the
Page 147 and 148: and HistoPARK (CZ.1.07/2.3.00/20.01
Page 149 and 150: Our results show that both p38α +/
Page 151 and 152: P90. TOLL-LIKE RECEPTOR 2 TRACKS TH
Page 153 and 154: for chemokines in attracting “inf
Page 155: in immune organs (Bursa of Fabricii
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ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.