MP68-01 THE LONG NON-CODING RNA HOTAIR MODULATES EXOSOME CONTENT AND FUNCTION IN BLADDER CANCER TUMOR PROGRESSION

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research IV1 Apr 2015MP68-01 THE LONG NON-CODING RNA HOTAIR MODULATES EXOSOME CONTENT AND FUNCTION IN BLADDER CANCER TUMOR PROGRESSION Claudia Berrondo, Jonathan Flax, Edward Messing, and Carla Beckham Claudia BerrondoClaudia Berrondo More articles by this author , Jonathan FlaxJonathan Flax More articles by this author , Edward MessingEdward Messing More articles by this author , and Carla BeckhamCarla Beckham More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.2463AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Exosomes are 30-100nM membrane-bound vesicles that participate in intercellular communication and alter tumor microenvironments. Exosomes deliver biologically active molecules like mRNA, protein, miRNA and long non-coding RNA (lncRNA) that affects the metabolism of recipient cells to produce pathological phenotypes. Exosomes may also activate the expression of genes involved in epithelial-to-mesenchyme transition (EMT) in recipient cells. Overexpression of the long non-coding RNA (lncRNA) HOTAIR (HA) is associated with poor prognosis and affects tumor progression, in part, by recruiting PRC2 and LSD1 chromatin repressive complexes to thousands of target genes. Objective: Identify if HA regulates exosome-mediated bladder cancer (BC) tumor progression. METHODS Tumor, distal normal tissue and urine from patients were obtained with IRB approval. HA levels were determined in patient samples, BC cell lines, and exosomes by qRT-PCR. An shRNA lentiviral vector was used to knockdown HA in T24 and TCC-SUP BC cells. Exosomes were harvested from shHOTAIR (shHA) and shScramble (shScr) BC cells. Migration and Invasion was evaluated by wound-healing, trans-well and 3-D culture assays, respectively. Changes in epithelial-to-mesenchyme transition (EMT) gene expression in exosome treated cells were measured by qRT-PCR. Coomassie gel of shScr and shHA BC cell exosomes was performed and bands representing differentially expressed proteins were sent for mass spectrometry. RESULTS HA is enriched in BC patient tumors, urinary exosome as well as BC cell lines. Knockdown of HA in BC cell lines significantly reduces migration and invasion in scratch, trans-well and 3-D culture assays; this correlates with altered expression of EMT genes. Exosomes isolated from shHA vs shScr cells fail to facilitate and in some cases inhibits migration of BC cell lines. A Coomassie gel of shScr and shHA exosomes revealed different relative levels of several proteins. CONCLUSIONS Loss of HA reduces migration and invasion of BC cells and these cells produce exosomes with reduced capacity to facilitate tumor progression. Our data is consistent with a model wherein loss of HA results in large-scale gene expression changes that ultimately affects exosome content. Taken together, these data suggest that exosomes could participate in and reflect the complexity of tumor heterogeneity as cells with particular genetic lesions produce exosomes with variable abilities to facilitate tumor progression. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e858 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Claudia Berrondo More articles by this author Jonathan Flax More articles by this author Edward Messing More articles by this author Carla Beckham More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...