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Profiling of microRNAs in exosomes released from PC-3 prostate cancer cells
Authors:Nina Pettersen Hessvik  Santosh Phuyal  Andreas Brech  Kirsten Sandvig  Alicia Llorente
Institution:1. Department of Biochemistry, Institute for Cancer Research, Oslo University Hospital–The Norwegian Radium Hospital, 0379 Oslo, Norway;2. Centre for Cancer Biomedicine, Faculty of Medicine, University of Oslo, 0379 Oslo, Norway;3. Department of Molecular Biosciences, University of Oslo, 0316 Oslo, Norway;1. Institut Curie, Centre de Recherche, 26 rue d’Ulm, Paris F-75248, France;2. INSERM U932, Paris F-75248, France;3. Paris Sciences et Lettres (PSL*), Paris F-75005, France;1. Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing 210009, China;2. Cardiovascular Disease, Internal Medicine, University of Cincinnati, 231 Albert Sabin Way, Cincinnati, OH 45267, United States;3. Internal Medicine of Traditional Chinese Medicine, Shuguang Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;4. Department of Cardiology, Xinhua Hospital, Shanghai Jiao Tong University, Shanghai, China;5. Feinberg Cardiovascular Research Institute, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, United States;6. Pathology and Lab Med, University of Cincinnati, 231 Albert Sabin Way, Cincinnati, OH 45267, United States;1. Center of Drug Metabolism and Pharmacokinetics, College of Pharmaceutical Sciences, Soochow University, Suzhou 215123, China;2. Department of Oncology, The Fourth Affiliated Hospital of Soochow University, Wuxi 214062, China;3. Department of Clinical Laboratory, The Fourth Affiliated Hospital of Soochow University, Wuxi 214062, China;4. Department of Gastroenterology, The First Affiliated Hospital of Soochow University, Suzhou 215006, China;5. Jiangsu Institute of Clinical Immunology, The First Affiliated Hospital of Soochow University, Suzhou 215006, China;1. Department of Human Genetics, Saarland University, Homburg/Saar, Germany;2. Department of Obstetrics and Gynecology, IVF and Andrology Laboratory, Saarland University, Homburg/Saar, Germany;1. Key Laboratory of Genomics and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China;2. University of Chinese Academy of Sciences, Beijing 100049, China;3. Department of Obstetrics and Gynecology, Jinan Central Hospital, Shandong University, Shandong 250013, China;4. State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China
Abstract:Exosomes are small extracellular vesicles released to the extracellular milieu through fusion of multivesicular bodies with the plasma membrane. These vesicles contain microRNAs and might therefore be vehicles transferring genetic information between cells. The aim of this study was to investigate whether there was a sorting of microRNAs into exosomes in the prostate cancer cell line PC-3. In addition, microRNAs in PC-3 cells and in the non-cancerous prostate cell line RWPE-1 were compared. Exosomes were isolated from the conditioned media from PC-3 cells by ultracentrifugation and inspected by electron microscopy. Total RNA was isolated and microRNAs were analyzed by microarray analysis and real time RT-PCR. MicroRNA microarray analysis revealed that the microRNA profile of PC-3 released exosomes was similar to the profile of the corresponding parent cells. Nevertheless, a sorting of certain microRNAs into exosomes was observed, and low number microRNAs (microRNAs with a low number in their name) were found to be underrepresented in these vesicles. Moreover, the miRNA profile of PC-3 cells resembled the miRNA profile of RWPE-1 cells, though some miRNAs were found to be differently expressed in these cell lines. These results show that exosomes from PC-3 cells, in agreement with previous reports from other cell types, contain microRNAs. Furthermore, this study supports the idea that there is a sorting of microRNAs into exosomes and adds a new perspective by pointing at the underrepresentation of low number miRNAs in PC-3 released exosomes.
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