239 MOLECULAR MECHANISMS OF SPECIFIC INHIBITION OF RENAL CELL CANCER (RCC) CELL MIGRATION BY THE DISINTEGRIN ECHISTATIN

Abstract

You have accessJournal of UrologyKidney Cancer: Basic Research1 Apr 2011239 MOLECULAR MECHANISMS OF SPECIFIC INHIBITION OF RENAL CELL CANCER (RCC) CELL MIGRATION BY THE DISINTEGRIN ECHISTATIN Wolfgang Jäger, Frederik Roos, Anna Becker, Elke Schneider, Joachim Thüroff, Christian Hampel, and Walburgis Brenner Wolfgang JägerWolfgang Jäger Mainz, Germany More articles by this author , Frederik RoosFrederik Roos Mainz, Germany More articles by this author , Anna BeckerAnna Becker Mainz, Germany More articles by this author , Elke SchneiderElke Schneider Mainz, Germany More articles by this author , Joachim ThüroffJoachim Thüroff Mainz, Germany More articles by this author , Christian HampelChristian Hampel Mainz, Germany More articles by this author , and Walburgis BrennerWalburgis Brenner Mainz, Germany More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.308AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Metastasis depends on adhesion of tumor cells to the endothelium, migration into subendothelial tissue and proliferation in secondary organs. Particularly during cell migration, binding of integrins, a group of cell adhesion molecules, to extracellular matrix is mediated by the amino acid sequence arginine-glycine-aspartate (RGD). Disintegrins, RGD containing proteins, can block integrins and thus inhibit cell migration and possibly metastasis. We analyzed the influence of disintegrins kistrin and echistatin on the migration of RCC cells. METHODS RCC-RC1 cells were pre-incubated with kistrin, echistatin and RGD containing peptides (P-RGD-M analogue to kistrin, A-RGD-D analogue to echistatin, G-RGD-S analogue to fibronectin, and RGD tripeptide). Migration was subsequently quantified in a boyden chemotaxis chamber. To analyse the binding mechanisms of these molecules, maleic anhydride activated plates (Pierce) were coated with echistatin, kistrin and the RGD containing molecules and the adherence of tumor cells, pre-incubated with á5- and 1-integrin-blocking antibodies, was quantified. The importance of the tertiary structure of the disintegrins was analysed by previous splitting of the disulfide chains. RESULTS In contrast to kistrin, echistatin significantly reduces tumor cell migration. Of the RGD containing peptides, G-RGD-S inhibited cell migration up to 45% and A-RGD-D up to 80%, whereas P-RGD-M did not show any inhibitory influence. Tumor cell adhesion was equally strong in echistatin and kistrin, and to a lower extent in fibronectin, but there was only a very weak binding to other RGD containing peptides. Previous incubation of RCC cells with integrin-blocking antibodies did not have any influence on tumor cell adhesion either. After splitting the disulfide chains of the disintegrins, the inhibitory effect of echistatin on tumor cell migration was reversed. Tumor cells also showed no adhesion to the modified echistatin. CONCLUSIONS Both disintegrins, echistatin and kistrin, contain the amino acid sequence RGD in equal amounts. After incubating RCC cells with disintegrins, only echistatin inhibited tumor cell migration. This effect seems not to be caused by a different binding capacity of these agents. After splitting the disulfide chains of echistatin and consequently destructing its proper tertiary structure, tumor cells lost adhesion capability. Subsequently inhibition of cell migration was not possible. Therefore a proper placement of the amino acid sequence RGD in echistatin is crucial for its inhibitory effects. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e97 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Wolfgang Jäger Mainz, Germany More articles by this author Frederik Roos Mainz, Germany More articles by this author Anna Becker Mainz, Germany More articles by this author Elke Schneider Mainz, Germany More articles by this author Joachim Thüroff Mainz, Germany More articles by this author Christian Hampel Mainz, Germany More articles by this author Walburgis Brenner Mainz, Germany More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...