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Session X : Cell death in cancer Poster X, 91 Functional analysis of the tumorsuppressor Ras Association Family Domain 1 (RASSF1A) reveals an important role of the SARAH-domain Claudia Seidel, Antje Klagge, Reinhard Dammann AWG Tumor Genetics of the Medical Faculty, Martin-Luther-University Halle- Wittenberg, 06097 Halle/Saale, Germany INTRODUCTION: RASSF1A is a tumor suppressor gene, which is often epigenetically inactivated in several cancer types. In lung cancer, a rate of between 30 to 45% of hypermethylation of the RASSF1A promoter has been found. In several cancer types a correlation between hypermethylation of RASSF1A and an impaired prognosis for cancer patients was reported. The promoter of the RASSF1C isoform is never hypermethylated. The two isoforms RASSF1A and RASSF1C only differ in the first domain, which is a diacylglycerol binding site and lacks in RASSF1C. Moreover RASSF1A encodes a Ras association domain and a SARAH domain, which is a protein-protein interaction domain. METHODS: In order to investigate the function of RASSF1A, we created mutations and deletions of these different domains and performed functional experiments in the lung cancer cell line A549. Proliferation analyses as well as soft agar experiments were performed to investigate the influence of the aberrant RASSF1A forms. The ability of the different forms of RASSF1A to induce apoptosis was investigated. To analyse the localisation of RASSF1A and its altered forms the cells were transient transfected using Yellow Fluorescent Protein (YFP) constructs. Additionally, the interaction of RASSF1A with the proapoptotic kinase MST1 was determined in precipitation experiments with protein extract from human cells. RESULTS: The reduced proliferation of A549 cells that express RASSF1A could be confirmed. Analysis of cells stable transfected with RASSF1A with a deletion of the SARAH domain (RASSF1ADelSARAH) showed a even lower proliferation rate in comparison to RASSF1A. A higher rate of apoptosis in cells with RASSF1ADelSARAH could be observed using transient transfected cells. RASSF1A is when overexpressed as a YFP-fusion colocalised to the cytoskeleton and during mitosis to the spindles and spindle poles. Using RASSF1ADelSARAH the mitoses show strong spindles no spindle poles and the chromosomes are not divided equally. In precipitation experiments MST1 interacts with RASSF1A but not with RASSF1ADelSARAH. CONCLUTIONS: The SARAH domain is necessary for the function of the tumor suppressor RASSF1A. It is responsible for the interaction with MST1 and plays an important role in regulation of mitotic processes as well as in induction of apoptosis. This study was supported by BMBF grant 01ZZ0104. - 424 -
Session X : Cell death in cancer Poster X, 92 ASSESSMENT OF EFFECT OF ECHINACEA PURPUREA ON APOPTOTIC AND MITOTIC ACTIVITY OF LIVER CELLS DURING INTOXICATION BY CADMIUM Alina Smalinskiene1, Stanislovas Ryselis1, Oleg Abdrakhmanov1, Rima Kregzdyte1, Vaiva Lesauskaite2, Ilona Sadauskiene1, Leonid Ivanov1, Nijole Savickiene3, Virgilijus Zitkevi(ius3, Arunas Savickas4 1Institute for Biomedical Research, Kaunas University of Medicine, 2Institute of Cardiology, Kaunas University of Medicine, 3Department of Pharmaceutical Chemistry and Pharmacognosy, Kaunas University of Medicine, 4Department of Drug Technology and Pharmaceutical Management, Kaunas University of Medicine, Kaunas, Lithuania E-mail : smalina@vector.kmu.lt Cadmium (Cd) is a very toxic and carcinogenic element. It disturbs the activity of biochemical systems of cells. Echinacea purpurea (L.) Moench (EP) can modify its influence. The aim of the study was to evaluate the effect of EP to accumulation of Cd in blood, liver, and kidney, mitotic and apoptotic activity of liver cells after the chronic intoxication by Cd2+. Materials and methods. Experiments were carried out on the white laboratory mice. Mice (n=57) were periodically i.p. injected for 6 weeks with CdCl2 (0.05 LD50 Cd2+) and EP extract solutions of two different concentrations (0.05 LD50 and 0.1 LD50) and their combinations. Control mice were injected with 0.9% saline. Cd concentration in blood and tissue specimens was determined by atomic absorption spectrophotometer Perkin- Elmer/Zeeman 3030. The number of mitotic liver cells was counted in 10 randomly selected reference areas (0.04 mm2). Apoptosis of liver cells was histochemically detected by the TUNEL assay using AP (Roche) in situ cell death detection kit. Preparation of extract from herb of Echinacea purpurea (L.) Moench was made according British herbal pharmacopoeia. Results. Cd2+ concentration in blood of mice in group EP(0.05)+Cd was 6.31-fold higher, therefore in group EP(0.1)+Cd was 8.25-fold higher comparing to Cd group, in liver that was 1.78 fold and 2.11-fold, respectively, in kidney 2.1-fold and 1.92-fold, respectively. Higher concentration (0.1LD50) of EP extract and Cd+EP 0.1LD50 arise apoptotic activity of liver cells to compare with control group. The mitotic activity of liver cells induced by Cd2+ after injection of EP extract was the same as in control group. Conclusion. Long-term injections of extract of EP (0.1 LD50) combined with CdCl2 (0.05 LD50) leads to the significant increase of cadmium concentration in blood and all investigated organs of experimental mice. EP decreases the mitotic activity of liver cells induced by cadmium and increases apoptotic activity of the liver cells. - 425 -
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Table of content PREFACE ..........
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Preface In 1998, we organized the f
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Acknowledgments This meeting has be
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Group B (15h00 - 17h00) Session V.
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- 11 - Exhibition
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Visit our Expo (in alphabetical ord
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Thursday January 26th, 2006 (Early
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Thursday January 26th, 2006 (Evenin
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Regulation of inflammation and gluc
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Saturday January 28th, 2006 (Early
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Oral presentations (by alphabetical
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4: : Lottin S, Vercoutter-Edouart A
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the B-Raf/mitogen-activated/extrace
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Albertini MC, Accorsi A, Citterio B
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AP-1-dependent gene expression in s
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Jaks and cytokine receptors - an in
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The Role of Met-Gab1 Signalling in
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The mammalian tumor suppressor Scri
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SIGNAL TRANSDUCTION BY STRESS-ACTIV
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Title to be announced Caroline Dive
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Mechanisms of DNA methylation in ma
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Blocking the "4 Integrin-Paxillin I
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Regulation of NF-kB-dependent trans
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The immunomodulator AS101 induces g
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Signaling pathways leading to the a
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Boute, N., Jockers, R. and Issad, T
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Jespsen JS, Sorensen MD and Wengel
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9) Proteome analysis of rat pancrea
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MicroRNA is an anti-apoptotic facto
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Resveratrol inhibits phorbol ester-
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Cross-talk between angiotensin II a
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Multiple role of histamine H 1 rece
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PI3K Targeting by the Beta-GBP Cyto
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D.P. Chopra, R.E. Menard, J. Janusz
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Oncogenic signaling by mutant p53 M
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activity of the Peroxisome Prolifer
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Restauration of SHIP-1 activity in
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Hypoxia Signaling. Impact on Tumor
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HOW ANTITOXIC AND ANTICANCER AGENTS
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Listeria monocytogenes induced Rac1
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Epigenetic Regulation of Stem Cell
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Survey on in vitro cell death induc
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Distinct roles of IRF-3 and IRF-7 i
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Redox-Sensitive Transcription Facto
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Epigenic control of MHC2TA transcri
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Attenuation of MSK1-driven NF-kB ge
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Talking to chromatin: Silencers Sil
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Ubiquitin ligase Smurf1 controls os
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Ambion MicroRNAs (miRNAs) are small
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Organizer: BIOBASE GmbH Date: Janua
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Reliable and efficient gene Knock-d
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Exploring ubiquitin signaling with
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Poster presentations Poster are cla
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Session I : protein domains Poster
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Session II. Receptor signaling and
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Session II : Receptor signaling and
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Session III : Protein kinase cascad
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IV. Protein kinase inhibitors: insi
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VI. Proteasome degradation pathways
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VI. Proteasome degradation pathways
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VIII. Inflammation specific signali
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Dr. Nassera Aouali L-1526 Luxembour
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