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

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Abstract

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.

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Acknowledgments

This study was ancillary to the PEAR study that was funded by the National Institutes of Health (U01 GM074492). Mohamed Hassan M. Solayman was funded by the Embassy of the Arab Republic of Egypt.

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Authors and Affiliations

  1. Department of Pharmacotherapy and Translational Research, Center for Pharmacogenomics, College of Pharmacy, University of Florida, Box 100484, Gainesville, FL, 32610-0484, USA

    Mohamed Hassan M. Solayman, Taimour Langaee, Archanakumari Patel & Julie A. Johnson

  2. Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt

    Mohamed Hassan M. Solayman, Lamia El-Wakeel, Manal El-Hamamsy & Osama Badary

Authors
  1. Mohamed Hassan M. Solayman
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  2. Taimour Langaee
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  3. Archanakumari Patel
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  4. Lamia El-Wakeel
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  5. Manal El-Hamamsy
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Correspondence to Julie A. Johnson.

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The authors declared no conflict of interest.

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All procedures were in accordance with the ethical standards of the corresponding institutional review boards and with the 1964 Helsinki declaration and its later amendments.

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Informed consent was obtained from all individual participants included in the study.

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Solayman, M.H.M., Langaee, T., Patel, A. et al. Identification of Suitable Endogenous Normalizers for qRT-PCR Analysis of Plasma microRNA Expression in Essential Hypertension. Mol Biotechnol 58, 179–187 (2016). https://doi.org/10.1007/s12033-015-9912-z

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  • DOI: https://doi.org/10.1007/s12033-015-9912-z

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