Identification of Suitable Endogenous Normalizers for qRT-PCR Analysis of Plasma microRNA Expression in Essential Hypertension

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• 作者：Mohamed Hassan M. Solayman ; Taimour Langaee ; Archanakumari Patel…
• 关键词：Essential hypertension ; Circulating miRNA ; Plasma ; Endogenous normalizer ; qRT ; PCR
• 刊名：Molecular Biotechnology
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：58
• 期：3
• 页码：179-187
• 全文大小：621 KB
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• 作者单位：Mohamed Hassan M. Solayman (1) (2)
  Taimour Langaee (1)
  Archanakumari Patel (1)
  Lamia El-Wakeel (2)
  Manal El-Hamamsy (2)
  Osama Badary (2)
  Julie A. Johnson (1)
  
  1. Department of Pharmacotherapy and Translational Research, Center for Pharmacogenomics, College of Pharmacy, University of Florida, Box 100484, Gainesville, FL, 32610-0484, USA
  2. Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt
  
• 刊物主题：Biotechnology; Biochemistry, general; Cell Biology; Protein Science; Biological Techniques; Human Genetics;
• 出版者：Springer US
• ISSN：1559-0305

文摘

Circulating microRNAs (miRNAs) are promising biomarkers for many diseases. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a gold standard for miRNA expression profiling that requires proper data normalization. Since there is no universal normalizer, it is recommended to evaluate normalizers under every experimental condition. This study describes the identification of suitable endogenous normalizer(s) (ENs) for plasma miRNA expression in essential hypertension. Expression levels of 5 candidate ENs and 2 plasma quality markers were determined by qRT-PCR in plasma samples from 18 hypertensive patients and 10 healthy controls. NormFinder, GeNorm, and DataAssist software programs were used to select the best EN(s). Expression levels of the 5 candidate ENs were also analyzed in urine samples from hypertensive patients and compared to the plasma samples of the hypertensive patients. Among the analyzed candidates, hsa-miR-92a-3p was identified as the best EN, and hsa-miR-21-5p and hsa-miR-16-5p as the next best. Moreover, hsa-miR-92a-3p showed the most consistent expression between plasma and urine. In conclusion, this study showed that hsa-miR-92a-3p, hsa-miR-21-5p, and hsa-miR-16-5p may be used as normalizers for plasma miRNA expression data in essential hypertension studies.