miRNA Normalization Candidates for qPCR in Lesional and Nonlesional Psoriatic Skin

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BACKGROUND: MicroRNA (miRNA) is an upcoming research area and quantitative reverse transcriptase in real-time (qPCR) is an important tool in the investigation. The small non-coding RNA candidates used for normalization in psoriatic skin biopsies have never been sufficiently validated. OBJECTIVES: To identify a reliable normalization method for miRNA analysis with qPCR in psoriatic skin. METHODS: 5 miRNA candidates previously used for normalization in psoriatic skin were identified through search in the literature. 5 new candidates were uncovered using the NormFinder algorithm on miRNA microArray data. The candidates were validated in paired psoriatic biopsies, biopsies from patients during treatment and normal healthy skin with qPCR. The stability of the miRNAs was determined with NormFinder and geNorm. The dispersion of data was determined before and after use of different normalization approaches. RESULTS: In lesional and nonlesional skin the two algorithms ranked the candidates similarly with an excellent correlation (R(2) =0.95). miR-26a had the best stability, whereas the commonly used RNU48 had less favourable stability. The same results were seen within the dispersion of the data, including biopsies from lesional, nonlesional, treated psoriatic skin and normal healthy skin. CONCLUSIONS: This is the first study to validate the reliability of miRNA candidates for normalization by qPCR in psoriatic skin. From this study we conclude that miR-26a is the best candidates and better than those previously used. miR-26a can be used for miRNA normalization in future studies with psoriatic skin. This article is protected by copyright. All rights reserved.
Original languageEnglish
JournalBritish Journal of Dermatology
ISSN0007-0963
DOIs
Publication statusPublished - 11 Apr 2013

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