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000127242 0247_ $$2doi$$a10.18632/oncotarget.6297
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000127242 037__ $$aDKFZ-2017-03267
000127242 041__ $$aeng
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000127242 1001_ $$aOleksiuk, Olga$$b0
000127242 245__ $$aSingle-Molecule Localization Microscopy allows for the analysis of cancer metastasis-specific miRNA distribution on the nanoscale.
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000127242 520__ $$aWe 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.
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000127242 650_7 $$2NLM Chemicals$$aMIRN31 microRNA, human
000127242 650_7 $$2NLM Chemicals$$aMicroRNAs
000127242 7001_ $$aAbba, Mohammed$$b1
000127242 7001_ $$aTezcan, Kerem Can$$b2
000127242 7001_ $$aSchaufler, Wladimir$$b3
000127242 7001_ $$aBestvater, Felix$$b4
000127242 7001_ $$aPatil, Nitin$$b5
000127242 7001_ $$aBirk, Udo$$b6
000127242 7001_ $$aHafner, Mathias$$b7
000127242 7001_ $$0P:(DE-He78)e6057131c71c90a6fcf61035f12b46c3$$aAltevogt, Peter$$b8$$udkfz
000127242 7001_ $$aCremer, Christoph$$b9
000127242 7001_ $$0P:(DE-He78)69067807288b48415ceb4abc43b9ad54$$aAllgayer, Heike$$b10$$eLast author$$udkfz
000127242 773__ $$0PERI:(DE-600)2560162-3$$a10.18632/oncotarget.6297$$gVol. 6, no. 42, p. 44745 - 44757$$n42$$p44745 - 44757$$tOncoTarget$$v6$$x1949-2553$$y2015
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000127242 9141_ $$y2015
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000127242 9201_ $$0I:(DE-He78)G360-20160331$$kG360$$lKKE Molekulare Onkologie solider Tumoren$$x1
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