TY  - JOUR
A1  - Köberle, Verena
A1  - Pleli, Thomas
A1  - Schmithals, Christian
A1  - Alonso, Eduardo Augusto
A1  - Haupenthal, Jörg
A1  - Bönig, Halvard-Björn
A1  - Peveling-Oberhag, Jan Franz-Josef
A1  - Biondi, Ricardo M.
A1  - Zeuzem, Stefan
A1  - Kronenberger, Bernd
A1  - Waidmann, Oliver
A1  - Piiper, Albrecht
T1  - Differential stability of cell-free circulating microRNAs : implications for their utilization as biomarkers
T2  - PLoS One
N2  - Background: MicroRNAs circulating in the blood, stabilized by complexation with proteins and/or additionally by encapsulation in lipid vesicles, are currently being evaluated as biomarkers. The consequences of their differential association with lipids/vesicles for their stability and use as biomarkers are largely unexplored and are subject of the present study.
Methods: The levels of a set of selected microRNAs were determined by quantitative reverse-transcription PCR after extraction from sera or vesicle- and non-vesicle fractions prepared from sera. The stability of these microRNAs after incubation with RNase A or RNase inhibitor, an inhibitor of RNase A family enzymes was studied.
Results: The levels of microRNA-1 and microRNA-122, but not those of microRNA-16, microRNA-21 and microRNA-142-3p, declined significantly during a 5-h incubation of the sera. RNase inhibitor prevented the loss of microRNAs in serum as well as the degradation of microRNA-122, a microRNA not expressed in blood cells, in whole blood. Stabilization of microRNA-122 was also achieved by hemolysis. Prolonged incubation of the sera led to enrichment of vesicle-associated relative to non-vesicle-associated microRNAs. Vesicle-associated microRNAs were more resistant to RNase A treatment than the respective microRNAs not associated with vesicles.
Conclusions: Serum microRNAs showed differential stability upon prolonged incubation. RNase inhibitor might be useful to robustly preserve the pattern of cell-free circulating microRNAs. In the case of microRNAs not expressed in blood cells this can also be achieved by hemolysis. Vesicle-associated microRNAs appeared to be more stable than those not associated with vesicles, which might be useful to disclose additional biomarker properties of miRNAs.
Y1  - 2013
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/31682
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-316828
SN  - 1932-6203
N1  - Copyright: © 2013 Köberle et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
VL  - 8
IS  - (9):e75184
PB  - PLoS
CY  - Lawrence, Kan.
ER  -