Protocol for analysis of miRNAs in human melanoma cells

Summary: MicroRNAs (miRNAs) significantly contribute to melanoma plasticity, a key factor in metastasis and therapy resistance. Here, we provide a protocol for analysis of miRNAs in human melanoma cells. We describe two approaches for plating cells, either using cell chambers or attaching cells to m...

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Main Authors: Evelyn Lattmann, Tessa Guggiari, Patrick Turko, Mitchell P. Levesque
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725002679
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author Evelyn Lattmann
Tessa Guggiari
Patrick Turko
Mitchell P. Levesque
author_facet Evelyn Lattmann
Tessa Guggiari
Patrick Turko
Mitchell P. Levesque
author_sort Evelyn Lattmann
collection DOAJ
description Summary: MicroRNAs (miRNAs) significantly contribute to melanoma plasticity, a key factor in metastasis and therapy resistance. Here, we provide a protocol for analysis of miRNAs in human melanoma cells. We describe two approaches for plating cells, either using cell chambers or attaching cells to microscopy slides by means of centrifugation. We first focus on cell pretreatment and staining procedures using the RNAscope Plus small RNA-RNA high-definition (RNAscope Plus smRNA-RNA HD) assay. We then detail the quantitative analysis of miRNA expression and co-localization. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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publishDate 2025-06-01
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spelling doaj-art-f22fd2f8b43242f5bdc53bfc7ea513f42025-08-20T02:00:52ZengElsevierSTAR Protocols2666-16672025-06-016210386110.1016/j.xpro.2025.103861Protocol for analysis of miRNAs in human melanoma cellsEvelyn Lattmann0Tessa Guggiari1Patrick Turko2Mitchell P. Levesque3Department of Dermatology, University of Zurich, University Hospital Zurich, 8952 Schlieren, Switzerland; Corresponding authorDepartment of Dermatology, University of Zurich, University Hospital Zurich, 8952 Schlieren, SwitzerlandDepartment of Dermatology, University of Zurich, University Hospital Zurich, 8952 Schlieren, SwitzerlandDepartment of Dermatology, University of Zurich, University Hospital Zurich, 8952 Schlieren, Switzerland; Corresponding authorSummary: MicroRNAs (miRNAs) significantly contribute to melanoma plasticity, a key factor in metastasis and therapy resistance. Here, we provide a protocol for analysis of miRNAs in human melanoma cells. We describe two approaches for plating cells, either using cell chambers or attaching cells to microscopy slides by means of centrifugation. We first focus on cell pretreatment and staining procedures using the RNAscope Plus small RNA-RNA high-definition (RNAscope Plus smRNA-RNA HD) assay. We then detail the quantitative analysis of miRNA expression and co-localization. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725002679Cell BiologyCell cultureCell-based AssaysCancerMicroscopyIn Situ Hybridization
spellingShingle Evelyn Lattmann
Tessa Guggiari
Patrick Turko
Mitchell P. Levesque
Protocol for analysis of miRNAs in human melanoma cells
STAR Protocols
Cell Biology
Cell culture
Cell-based Assays
Cancer
Microscopy
In Situ Hybridization
title Protocol for analysis of miRNAs in human melanoma cells
title_full Protocol for analysis of miRNAs in human melanoma cells
title_fullStr Protocol for analysis of miRNAs in human melanoma cells
title_full_unstemmed Protocol for analysis of miRNAs in human melanoma cells
title_short Protocol for analysis of miRNAs in human melanoma cells
title_sort protocol for analysis of mirnas in human melanoma cells
topic Cell Biology
Cell culture
Cell-based Assays
Cancer
Microscopy
In Situ Hybridization
url http://www.sciencedirect.com/science/article/pii/S2666166725002679
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