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Session X : Cell death in cancer Poster X, 61 HGF/SF regulates expression of apoptotic genes in human mammary epithelial cells Catherine LEROY, Julien DEHEUNINCK, Sylvie REVENEAU, Bénédicte FOVEAU, Zongling JI, David TULASNE, Céline Villenet*, Sabine Quief*, and Jean-Pierre Kerckaert* and Véronique FAFEUR. CNRS UMR 8117, Institut de Biologie de Lille, Institut Pasteur de Lille, B.P.447, 59021 Lille, FRANCE. E-mail : veronique.fafeur@ibl.fr * Plate-forme Biopuces, Université de Lille 2, Faculté de Médecine, Place de Verdun, 59045, Lille, FRANCE Hepatocyte growth factor/scatter factor (HGF/SF) induces scattering, morphogenesis and survival of epithelial cells through the activation of the MET tyrosine kinase receptor. HGF/SF and MET are involved in normal and tumoral development of many tissues and organs, including the mammary gland. At present, little is known of the target genes of HGF/SF implicated in mediating its biological responses. We searched for HGF/SF target genes using the human epithelial mammary cell line (MCF- 10A), which is sensitive to its scattering and survival effect. We used a DNA microarray spotted with 60-mer oligonucleotide sets from Sigma-Genosys that include 1920 different genes. MCF-10A cells were treated or not with HGF/SF for 2 h. Total RNA was then extracted and purified, followed by reverse transcription to cDNA, with concomitant incorporation of fluorescent dCTP (Cy3 or Cy5). The probes were mixed and hybridized onto the microarray, and the fluorescent signals were detected using a GMS confocal scanner (Affymetrix). The ratios of Cy5/Cy3 were analyzed using Jaguar Software (Screensaver). The expression of 38 genes was found to be modified by HGF/SF. Among them, only three genes were clearly classified in apoptosis, with A20 being up-regulated by HGF/SF and DAXX and SMAC being down-regulated by HGF/SF. Changes in expression of A20, DAXX and SMAC were confirmed by real time quantitative PCR using a Light Cycler (Roche). According to published data, A20 is anti-apoptotic, SMAC is pro-apoptotic, while a pro- or anti- apoptotic role of DAXX is still controversial. The fact that HGF/SF up-regulates an anti-apoptotic gene (A20) and down-regulates a pro-apoptotic gene (SMAC) is in agreement with its survival effect in MCF10A cells. This study identified novel target genes of HGF/SF that can contribute to its anti-apoptotic effect. - 394 -
Session X : Cell death in cancer Poster X, 62 The nucleoside analogue cidofovir suppresses lung metastasis and induces apoptosis in FGF2-overexpressing endothelial cells. Sandra Liekens*, Sofie Gijsbers*, Erik De Clercq*, Erik Verbeken§, and Sigrid Hatse* *Rega Institute for Medical Research, Minderbroedersstraat 10, B-3000 Leuven, Belgium and §Division of Histopathology, K.U.Leuven. E-mail: Sandra.liekens@rega.kuleuven.be Cidofovir [(S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine, (S)-HPMPC] is an antiviral drug that is clinically used for the treatment of cytomegalovirus retinitis in AIDS patients (1). Cidofovir also possesses potent activity against human papillomavirus-induced tumors in animal models and patients (1). We have recently shown that cidofovir inhibits the development of vascular tumors induced by basic fibroblast growth factor (FGF2)overexpressing endothelial (3F2T) cells in mice (2). Here, we demonstrate that the cytotoxic activity of cidofovir in 3F2T cells may result from the specific induction of apoptosis. Cell cycle analysis revealed that cidofovir induces accumulation of cells in the S phase and, upon prolonged treatment, a significant increase in sub-G1 cells, exhibiting a sub-diploid DNA content. Moreover, annexin V binding, an early event in apoptosis induction, was increased in cidofovir-treated 3F2T cells. Cidofovir also caused nuclear fragmentation and the activation of caspase-3-like proteases in 3F2T cells, as evidenced by the cleavage of poly(ADPribose)polymerase. In addition, cidofovir treatment resulted in a pronounced up-regulation of p53 protein. Protein kinase B/Akt is recognized as a principal mediator of survival signals that protect cells from undergoing apoptosis. Cidofovir did not suppress the phosphorylation of Akt nor its downstream regulator Bad, indicating that the Akt pathway is not affected by cidofovir treatment of 3F2T cells. However, cidofovir inhibited the expression of FGF2 and FGF2 signaling through Erk42/44, as shown by Western blot analysis. In vivo, cidofovir markedly suppressed the development of experimental lung metastasis, induced by inoculation of 3F2T cells in the tail vein of SCID mice. Our results indicate that cidofovir may inhibit the growth of 3F2T cells via inhibition of FGF2 expression and signaling, and via the induction of apoptosis through the caspase-3 pathway. 1. De Clercq and Holy (2005) Nat Rev Drug Discov. 4: 928-940. 2. Liekens et al. (2001) Cancer Res. 61: 5057-5064. - 395 -
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- 1 - Luxembourg, January 25th to 2
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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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8. Nishikawa, H., Kawai, K., Tanaka
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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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[2] Meijer, L., Flajolet, M. and Gr
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[4] Niemand, C., Nimmesgern, A., Ha
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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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5. Sokolov EI, Zykov KA, Pukhalsky
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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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Workshop presentations (by alphabet
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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 cascade
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Session III : Protein kinase cascad
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Session IV. Protein kinase inhibito
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IV. Protein kinase inhibitors: insi
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IV. Protein kinase inhibitors: insi
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V. Phosphatases as key cell signali
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VI. Proteasome degradation pathways
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VI. Proteasome degradation pathways
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Session VII. Stem cell specific cel
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VIII. Inflammation specific signali
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Session IX. Novel compounds targeti
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Session X : Cell death in cancer -
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Session X : Cell death in cancer Po
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Session XI : Cell death and cardiov
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Session XI: Cell death and cardiova
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Session XII : Cell death and neurod
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Session XIII : Cell signaling pathw
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Session XIV : Transcriptional and t
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Session XV : Reactive oxygen specie
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Session XVI : Chemopreventive agent
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Session XVII : Cell signaling in he
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Dr. Nassera Aouali L-1526 Luxembour
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Dr. Giordano Bianchi Clinic of inte
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Mrs. Isabel Burghardt Laboratory of
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Pr. Czeslaw Cierniewski 92-215 Lodz
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Dr. Mark Ginsberg Mail code 0726 92
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Dr. Jochen Hefl Division of Signal
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Mr. Jan Jepsen 2100 Copenhagen DNK
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Pr. Young Min Kim Division of Biolo
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Mr. Christoph Lahtz AG Tumorgenetik
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Mrs. Rasa Merzvinskyte Department o
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Mr. Gustav Nilsonne Department of L
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Mrs. Christel Péqueux Tumour and D
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Dr. Gabriella Regis Tumor Immunolog
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