MP49-07 CANONICAL WNT PATHWAY TUMOR PROGRESSION IS MEDIATED BY THE LONG NON-CODING RNA HOTAIR IN BLADDER CANCER

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research III1 Apr 2015MP49-07 CANONICAL WNT PATHWAY TUMOR PROGRESSION IS MEDIATED BY THE LONG NON-CODING RNA HOTAIR IN BLADDER CANCER 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.2015.02.511AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Overexpression of the long non-coding RNA (lncRNA) HOTAIR (HA) is associated with poor prognosis and affects tumor progression, in part, by recruiting chromatin repressive complexes PRC2 and LSD1 to thousands of target genes including antagonists of Wnt-mediated EMT. Recently the BC Genomic Atlas Project identified that 67% of BC tumors have some perturbation in the canonical Wnt pathway. Importantly, elevated levels of HA were associated with BC tumor recurrence. Here we show that HA levels increase in response to activation of the Wnt pathway and HA expression is necessary for migration in response to Wnt3a stimulation. Objective: Identify if HA is a target of and participant in the canonical Wnt pathway. METHODS Tumor and distal normal tissue was collected from BC patients with IRB approval. HA levels in patient samples and BC cell lines: 5637, T24, and TCC-SUP was determined by qRT-PCR. shRNA lentiviral vectors were used to knockdown HA in T24 and TCC-UP cells. Migration and Invasion was evaluated via scratch, trans-well and 3-D culture, respectively. shScr and shHA cells were treated with LiCl (a non-specific activator of the Wnt pathway) or recombinant Wnt3a (rWnt3a). Activation of the Wnt pathway was assessed by qRT-PCR for expression of Wnt target genes. Migration was evaluated by scratch assay. DNA sequencing of HA promoter region for TCF7L2 (TCF4) binding sites was performed in several BC cell lines. RESULTS HA is enriched in BC patient tumors and BC cell lines. Treatment of BC cell with either LiCl or rWnt3a led to increased levels of HA and Wnt target genes: AXIN2, CCND1 and ZEB1. shHA cells show significantly reduced migration and invasion in trans-well and 3-D culture. rWnt3a treatment of BC cells increases migration, while shHA cells fail to migrate in response to rWnt3a. The expression of several genes involved in migration and EMT (SNAIL, LAMC2, LAMB3, ZO, and ZEB1) are altered in shHA knockdown BC cells. CONCLUSIONS HA is a mediator of tumor progression in several solid tumors. We show that HA mediates responsiveness to activation of the canonical Wnt pathway in BC possibly through mediating expression of Wnt-associated EMT factors. HA levels increase in response to Wnt pathway stimulation. Wnt pathway activation results in transcription of target genes via β-catenin binding to TCF7L2 sites. We show BC cells contain TCF7L2 sites, suggesting HA maybe a downstream target of the Wnt pathway. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e605 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 ...