MP45-01 EXOSOMAL MIRNAS: KEY REGULATORS OF CELL-CELL COMMUNICATION BETWEEN BLADDER CANCER CELLS AND TUMOR MICROENVIRONMENT

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology I1 Apr 2016MP45-01 EXOSOMAL MIRNAS: KEY REGULATORS OF CELL-CELL COMMUNICATION BETWEEN BLADDER CANCER CELLS AND TUMOR MICROENVIRONMENT Sophie Baumgart, Joana Heinzelmann, Elmar Krause, Marie Stampe Ostenfeld, Arndt Hartmann, 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 , Arndt HartmannArndt Hartmann 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.2016.02.278AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Interaction of tumor cells and tumor microenvironment (TME) plays an important role in tumor progression. It was shown that microRNAs (miRNAs) packed in exosomes (EV) can affect cell-cell communication at the site of origin (TME) as well as distant sites. One aim of the project is the identification of a specific miRNA expression pattern from tumor-derived EV of different urinary bladder cancer (UBC) cell lines in correlation to their invasive potential. Afterwards, the functional role of specific exosomal miRNAs on cells of the TME (e.g. naive fibroblasts) will be analyzed. METHODS EV were isolated from invasive (T-24,253J-BV, J82) and non-invasive (RT-112, 5637) UBC cell lines and characterized by Western Blot analysis, Nano Particle Tracking Analysis and electron microscopy. MiRNA expression analysis in UBC cells and their EV was performed by using miRNA microarray and qPCR. Uptake of PKH26-labeled UBC-EV by immortalized fibroblasts was shown by laser scanning microscopy. EV-mediated miRNA transfer between cancer cells and fibroblasts was verified by transfection of donor UBC cells with the C. elegans-specific miRNA, cel-miR-39, EV isolation and RNAse treatment, transfer to recipient fibroblasts (TAFs and naive fibroblasts), and miRNA-specific qPCR analysis using totalRNA from the recipient fibroblasts. RESULTS Electron microscopy images revealed the typical size and vesicular structure of EV. The size measurement revealed a mean size of EV among 61-81nm. Isolated EV exhibited a high amount of exosomal markers (CD81, syntenin) and less presence of contamination marker calreticulin. 16 miRNAs were identified distinguishing invasive UBC cells from non-invasive cells. EV secreted by invasive UBC cells are characterized by a specific miRNA signature of 25 miRNAs compared to EV from non-invasive UBC cells. 6 significantly different expressed miRNAs clearly separate both invasive and non-invasive UBC cells and their secreted EV. Labeled EV of UBC cells were taken up by naive fibroblasts. After co-cultivation of EV obtained from transfected UBC cells and naive fibroblasts, high expression of cel-miR-39 was detected in naive fibroblasts. CONCLUSIONS EV secreted by UBC cells exhibit a specific miRNA signature depending on the invasive potential of the originating cells. For the first time we demonstrated the EV-mediated transfer of miRNAs between UBC cells and naive fibroblasts. Furthermore, EV of UBC cells can be taken up by fibroblasts. These results emphasize the role of exosomal miRNAs for the interaction between tumor cells and the tumor microenvironment. Further studies have to show the functional relevance of selected exosomal miRNAs. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e607 Advertisement Copyright & Permissions© 2016MetricsAuthor 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 Arndt Hartmann 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 ...