TY  - JOUR
AU  - Oleksiuk, Olga
AU  - Abba, Mohammed
AU  - Tezcan, Kerem Can
AU  - Schaufler, Wladimir
AU  - Bestvater, Felix
AU  - Patil, Nitin
AU  - Birk, Udo
AU  - Hafner, Mathias
AU  - Altevogt, Peter
AU  - Cremer, Christoph
AU  - Allgayer, Heike
TI  - Single-Molecule Localization Microscopy allows for the analysis of cancer metastasis-specific miRNA distribution on the nanoscale.
JO  - OncoTarget
VL  - 6
IS  - 42
SN  - 1949-2553
CY  - [S.l.]
PB  - Impact Journals LLC
M1  - DKFZ-2017-03267
SP  - 44745 - 44757
PY  - 2015
AB  - We describe a novel approach for the detection of small non-coding RNAs in single cells by Single-Molecule Localization Microscopy (SMLM). We used a modified SMLM-setup and applied this instrument in a first proof-of-principle concept to human cancer cell lines. Our method is able to visualize single microRNA (miR)-molecules in fixed cells with a localization accuracy of 10-15 nm, and is able to quantify and analyse clustering and localization in particular subcellular sites, including exosomes. We compared the metastasis-site derived (SW620) and primary site derived (SW480) human colorectal cancer (CRC) cell lines, and (as a proof of principle) evaluated the metastasis relevant miR-31 as a first example. We observed that the subcellular distribution of miR-31 molecules in both cell lines was very heterogeneous with the largest subpopulation of optically acquired weakly metastatic cells characterized by a low number of miR-31 molecules, as opposed to a significantly higher number in the majority of the highly metastatic cells. Furthermore, the highly metastatic cells had significantly more miR-31-molecules in the extracellular space, which were visualized to co-localize with exosomes in significantly higher numbers. From this study, we conclude that miRs are not only aberrantly expressed and regulated, but also differentially compartmentalized in cells with different metastatic potential. Taken together, this novel approach, by providing single molecule images of miRNAs in cellulo can be used as a powerful supplementary tool in the analysis of miRNA function and behaviour and has far reaching potential in defining metastasis-critical subpopulations within a given heterogeneous cancer cell population.
KW  - MIRN31 microRNA, human (NLM Chemicals)
KW  - MicroRNAs (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:26561203
C2  - pmc:PMC4792589
DO  - DOI:10.18632/oncotarget.6297
UR  - https://inrepo02.dkfz.de/record/127242
ER  -