Different gDNA content in the subpopulations of prostate cancer extracellular vesicles: Apoptotic bodies, microvesicles, and exosomes

BACKGROUND Extracellular vesicles (EVs) are cell‐derived membrane vesicles. EVs contain several RNAs such as mRNA, microRNAs, and ncRNAs, but less is known of their genomic DNA (gDNA) content. It is also unknown whether the DNA cargo is randomly sorted or if it is systematically packed into specific...

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Published in:The Prostate Vol. 74; no. 14; pp. 1379 - 1390
Main Authors: Lázaro-Ibáñez, Elisa, Sanz-Garcia, Andres, Visakorpi, Tapio, Escobedo-Lucea, Carmen, Siljander, Pia, Ayuso-Sacido, Ángel, Yliperttula, Marjo
Format: Journal Article
Language:English
Published: United States Blackwell Publishing Ltd 01.10.2014
Wiley Subscription Services, Inc
BlackWell Publishing Ltd
Subjects:
Apoptosis - genetics
Case-Control Studies
Cell Line, Tumor
DNA, Neoplasm - genetics
DNA, Neoplasm - metabolism
Exosomes - genetics
Exosomes - metabolism
Genes, p53
Humans
Male
Original
Prostatic Neoplasms - blood
Prostatic Neoplasms - genetics
Prostatic Neoplasms - metabolism
Prostatic Neoplasms - pathology
PTEN Phosphohydrolase - genetics
ISSN:0270-4137, 1097-0045, 1097-0045
Online Access:Get full text
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Summary:BACKGROUND Extracellular vesicles (EVs) are cell‐derived membrane vesicles. EVs contain several RNAs such as mRNA, microRNAs, and ncRNAs, but less is known of their genomic DNA (gDNA) content. It is also unknown whether the DNA cargo is randomly sorted or if it is systematically packed into specific EV subpopulations. The aim of this study was to analyze whether different prostate cancer (PCa) cell‐derived EV subpopulations (apoptotic bodies, microvesicles, and exosomes) carry different gDNA fragments. METHODS EV subpopulations were isolated from three PCa cell lines (LNCaP, PC‐3, and RC92a/hTERT) and the plasma of PCa patients and healthy donors, and characterized by transmission electron microscopy, nanoparticle tracking analysis and total protein content. gDNA fragments of different genes were detected by real time quantitative PCR and confirmed by DNA sequencing. RESULTS We report that the concentration of EVs was higher in the cancer patients than in the healthy controls. EV subpopulations differed from each other in terms of total protein and DNA content. Analysis of gDNA fragments of MLH1, PTEN, and TP53 genes from the PCa cell‐derived EV subpopulations showed that different EVs carried different gDNA content, which could even harbor specific mutations. Altogether, these results suggest that both nucleic acids and proteins are selectively and cell‐dependently packed into the EV subtypes. CONCLUSIONS EVs derived from PCa cell lines and human plasma samples contain double‐stranded gDNA fragments which could be used to detect specific mutations, making EVs potential biomarkers for cancer diagnostics and prognostics. Prostate 74:1379–1390, 2014. © 2014 The Authors. The Prostate published by Wiley Periodicals, Inc.
Bibliography:ArticleID:PROS22853
Academy of Finland - No. 259990
Academy of Finland Research Fellow - No. 273689-266486
Finnish Cultural Foundation - No. 00130502
ark:/67375/WNG-F3P7WGZ9-G
Carlos III Health Institute - No. PI10/01069; No. CP11/00147
istex:C7B7F9FEA495F8F88F1860A685EBCA77312FA9A2
Magnus Ehrnrooth Foundation
Medicinska Understödsföreningen Liv och Hälsa r.f.
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SourceType-Scholarly Journals-1
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Grant sponsor: Finnish Cultural Foundation; Grant number: 00130502; Grant sponsor: Academy of Finland Research Fellow; Grant number: 273689–266486; Grant sponsor: Magnus Ehrnrooth Foundation; Grant sponsor: Medicinska Understödsföreningen Liv och Hälsa r.f.; Grant sponsor: Carlos III Health Institute; Grant numbers: PI10/01069; CP11/00147; Grant sponsor: Academy of Finland; Grant number: 259990.
ISSN:0270-4137
1097-0045
1097-0045
DOI:10.1002/pros.22853