MP54-15 UROTHELIAL BLADDER CANCER CELLS AFFECT TUMOR-PROMOTING PROCESSES IN NORMAL BLADDER FIBROBLASTS AND SUPPORT TUMORIGENESIS BY SECRETION OF TUMOR-ASSOCIATED EXOSOMES

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology I1 Apr 2018MP54-15 UROTHELIAL BLADDER CANCER CELLS AFFECT TUMOR-PROMOTING PROCESSES IN NORMAL BLADDER FIBROBLASTS AND SUPPORT TUMORIGENESIS BY SECRETION OF TUMOR-ASSOCIATED EXOSOMES Sophie Baumgart, Joana Heinzelmann, Elmar Krause, Marie Stampe-Ostenfeld, Michael Stöckle, and Kerstin Junker Sophie BaumgartSophie Baumgart More articles by this author , Joana HeinzelmannJoana Heinzelmann More articles by this author , Elmar KrauseElmar Krause More articles by this author , Marie Stampe-OstenfeldMarie Stampe-Ostenfeld More articles by this author , Michael StöckleMichael Stöckle More articles by this author , and Kerstin JunkerKerstin Junker More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1706AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES MiRNAs packed in exosomes (EV) can affect cell-cell communication in the tumor microenvironment and play a crucial role in tumorigenesis. Our aim is the identification of a specific exosomal miRNA expression pattern depending on the invasiveness of urinary bladder cancer (BC) cells. Furthermore we want to characterize their functional role in reprogramming normal bladder fibroblasts (BF). METHODS EVs were isolated from invasive (T24,253J-BV,J82), non-invasive (RT112,5637) BC cells and normal bladder cells (HCV29) by ultracentrifugation and were characterized by Western Blot analysis, Nano Particle Tracking Analysis and electron microscopy. Uptake of PKH26-labeled BC-EV by hTERT-immortalized fibroblasts (hTERT-FB) was shown by LSM. EV-mediated miRNA transfer between BC cells and hTERT-FB was analyzed by transfection of BC cells with cel-miR-39, co-cultivation with hTERT-FB and qPCR. Proliferation and migration of EV-treated normal BF were measured by BrdU assay and Boyden Chamber assay. RESULTS Electron microscopy images revealed the typical size (61-81 nm) and vesicular structure of EV. Isolated EV exhibited a high amount of exosomal markers (CD81, syntenin) and less presence of contamination marker calreticulin. EV of invasive BC cells are characterized by a specific miRNA signature of 15 miRNAs. Validation of 6 selected miRNAs confirms a significant different expression between both, invasive and non-invasive cells and their EV. miR-30a-3p could be confirmed by qPCR to be significantly deregulated exclusively in EVs of invasive cells compared to the non-invasive counterparts, but not in the parental cells. Uptake of labeled EV and exosomal miRNA of BC cells in hTERT-FB could be confirmed. Co-cultivation of tumor-associated and normal EV and normal BF resulted in increased proliferation and migration rates in normal BF independently from the invasiveness of donor BC cells, but cell-type specific. CONCLUSIONS EVs secreted by BC cells are characterized by a specific miRNA pattern depending on their invasiveness. We demonstrated that tumor-derived EV can stimulate the proliferation and migration of BF indicating an important role in cell-cell communication. Further studies have to investigate the changes of signaling pathways in fibroblasts induced by EV and their miRNAs and the role in the development of an invasive phenotype. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e717-e718 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Sophie Baumgart More articles by this author Joana Heinzelmann More articles by this author Elmar Krause More articles by this author Marie Stampe-Ostenfeld More articles by this author Michael Stöckle More articles by this author Kerstin Junker More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...