Endogenous reference RNAs for microRNA quantitation in formalin-fixed, paraffin-embedded lymph node tissue

Lymph node metastasis is one of the most important factors for tumor dissemination. Quantifying microRNA (miRNA) expression using real-time PCR in formalin-fixed, paraffin-embedded (FFPE) lymph node can provide valuable information regarding the biological research for cancer metastasis. However, a...

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Bibliographic Details
Published in:Scientific reports 2018-04, Vol.8 (1), p.5918-10, Article 5918
Main Authors: Inada, Katsushige, Okoshi, Yasushi, Cho-Isoda, Yukiko, Ishiguro, Shingo, Suzuki, Hisashi, Oki, Akinori, Tamaki, Yoshio, Shimazui, Toru, Saito, Hitoaki, Hori, Mitsuo, Iijima, Tatsuo, Kojima, Hiroshi
Format: Article
Language:English
Subjects:
38
38/77
631/1647/2017/2003
631/337/384/331
631/67/322
Biological research
Biopsy
Cancer
Formaldehyde
Genes
Humanities and Social Sciences
Lymph nodes
Lymphatic system
Metastases
Metastasis
MicroRNAs
miRNA
multidisciplinary
Paraffin
Quantitation
Science
Science (multidisciplinary)
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Summary:Lymph node metastasis is one of the most important factors for tumor dissemination. Quantifying microRNA (miRNA) expression using real-time PCR in formalin-fixed, paraffin-embedded (FFPE) lymph node can provide valuable information regarding the biological research for cancer metastasis. However, a universal endogenous reference gene has not been identified in FFPE lymph node. This study aimed to identify suitable endogenous reference genes for miRNA expression analysis in FFPE lymph node. FFPE lymph nodes were obtained from 41 metastatic cancer and from 16 non-cancerous tissues. We selected 10 miRNAs as endogenous reference gene candidates using the global mean method. The stability of candidate genes was assessed by the following four statistical tools: BestKeeper, geNorm, NormFinder, and the comparative ΔCt method. miR-103a was the most stable gene among candidate genes. However, the use of a single miR-103a was not recommended because its stability value exceeded the reference value. Thus, we combined stable genes and investigated the stability and the effect of gene normalization. The combination of miR-24, miR-103a, and let-7a was identified as one of the most stable sets of endogenous reference genes for normalization in FFPE lymph node. This study may provide a basis for miRNA expression analysis in FFPE lymph node tissue.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-018-24338-7