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Article

Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion

  • Authors:
    • Hye‑Suk Han
    • Yeong Nang Jo
    • Jin Yong Lee
    • Song‑Yi Choi
    • Yusook Jeong
    • Jieun Yun
    • Ok‑Jun Lee
  • View Affiliations / Copyright

    Affiliations: Department of Internal Medicine, College of Medicine, Chungbuk National University, Cheongju 361‑763, Republic of Korea, Bioevaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongwon 363‑883, Republic of Korea, Public Health Medical Service, SMG‑SNU Boramae Medical Center, Seoul 156‑707, Republic of Korea, Department of Pathology, College of Medicine, Chungbuk National University, Cheongju 361‑763, Republic of Korea
  • Pages: 1889-1895
    |
    Published online on: August 1, 2014
       https://doi.org/10.3892/ol.2014.2404
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Abstract

Circulating cell‑free microRNAs (miRNAs) are potential biomarkers of cancer. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) is widely used in miRNA expression studies. The aim of this study was to identify suitable reference genes for RT‑qPCR analyses of miRNA expression levels in pleural effusion. The expression levels of candidate reference miRNAs were investigated in 10 benign pleural effusion (BPE) and 10 lung adenocarcinoma‑associated malignant pleural effusion (LA‑MPE) samples using miRNA microarrays. The expression levels of candidate reference miRNAs, together with those of U6 small nuclear RNA (snRNA), RNU6B, RNU44 and RNU48 small RNAs, in 46 BPE and 45 LA‑MPE samples were validated by RT‑qPCR, and were analyzed using the NormFinder and BestKeeper algorithms. The impact of different normalization approaches on the detection of differential expression levels of miR‑198 in BPE and LA‑MPE samples was also assessed. As determined by the miRNA microarray data, five candidate reference miRNAs were identified. Following RT‑qPCR validation, U6 snRNA, miR‑192, miR‑20a, miR‑221, miR‑222 and miR‑16 were evaluated using the NormFinder and BestKeeper software programs. U6 snRNA and miR‑192 were identified as single reference genes and the combination of these genes was preferred for the relative quantification of miRNA expression levels in pleural effusion. Normalization of miR‑98 expression levels to those of U6 snRNA, miR‑192 or a combination of these genes enabled the detection of a significant difference between BPE and LA‑MPE samples. Therefore, U6 snRNA and miR‑192 are recommended as reference genes for the relative quantification of miRNA expression levels in pleural effusion.
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Copy and paste a formatted citation
Spandidos Publications style
Han HS, Jo YN, Lee JY, Choi SY, Jeong Y, Yun J and Lee OJ: Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion. Oncol Lett 8: 1889-1895, 2014.
APA
Han, H., Jo, Y.N., Lee, J.Y., Choi, S., Jeong, Y., Yun, J., & Lee, O. (2014). Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion. Oncology Letters, 8, 1889-1895. https://doi.org/10.3892/ol.2014.2404
MLA
Han, H., Jo, Y. N., Lee, J. Y., Choi, S., Jeong, Y., Yun, J., Lee, O."Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion". Oncology Letters 8.4 (2014): 1889-1895.
Chicago
Han, H., Jo, Y. N., Lee, J. Y., Choi, S., Jeong, Y., Yun, J., Lee, O."Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion". Oncology Letters 8, no. 4 (2014): 1889-1895. https://doi.org/10.3892/ol.2014.2404
Copy and paste a formatted citation
x
Spandidos Publications style
Han HS, Jo YN, Lee JY, Choi SY, Jeong Y, Yun J and Lee OJ: Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion. Oncol Lett 8: 1889-1895, 2014.
APA
Han, H., Jo, Y.N., Lee, J.Y., Choi, S., Jeong, Y., Yun, J., & Lee, O. (2014). Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion. Oncology Letters, 8, 1889-1895. https://doi.org/10.3892/ol.2014.2404
MLA
Han, H., Jo, Y. N., Lee, J. Y., Choi, S., Jeong, Y., Yun, J., Lee, O."Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion". Oncology Letters 8.4 (2014): 1889-1895.
Chicago
Han, H., Jo, Y. N., Lee, J. Y., Choi, S., Jeong, Y., Yun, J., Lee, O."Identification of suitable reference genes for the relative quantification of microRNAs in pleural effusion". Oncology Letters 8, no. 4 (2014): 1889-1895. https://doi.org/10.3892/ol.2014.2404
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