title: Identification of Suitable Endogenous Normalizers for qRT-PCR Analysis of Plasma microRNA Expression in Essential Hypertension.
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.
authors.affiliation: Department of Pharmacotherapy and Translational Research, Center for Pharmacogenomics, College of Pharmacy, University of Florida, Box 100484, Gainesville, FL, 32610-0484, USA.|Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
authors.forename: Mohamed Hassan M
authors.initials: MH
authors.lastname: Solayman
authors.affiliation: Department of Pharmacotherapy and Translational Research, Center for Pharmacogenomics, College of Pharmacy, University of Florida, Box 100484, Gainesville, FL, 32610-0484, USA.
authors.forename: Taimour
authors.initials: T
authors.lastname: Langaee
authors.affiliation: Department of Pharmacotherapy and Translational Research, Center for Pharmacogenomics, College of Pharmacy, University of Florida, Box 100484, Gainesville, FL, 32610-0484, USA.
authors.forename: Archanakumari
authors.initials: A
authors.lastname: Patel
authors.affiliation: Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
authors.forename: Lamia
authors.initials: L
authors.lastname: El-Wakeel
authors.affiliation: Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
authors.forename: Manal
authors.initials: M
authors.lastname: El-Hamamsy
authors.affiliation: Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
authors.forename: Osama
authors.initials: O
authors.lastname: Badary
authors.affiliation: Department of Pharmacotherapy and Translational Research, Center for Pharmacogenomics, College of Pharmacy, University of Florida, Box 100484, Gainesville, FL, 32610-0484, USA. johnson@cop.ufl.edu.
authors.forename: Julie A
authors.initials: JA
authors.lastname: Johnson
mesh_terms: D000328:Adult; D000075222:Essential Hypertension; D005260:Female; D020869:Gene Expression Profiling / Q000592:standards*; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease; D006801:Humans; D006973:Hypertension / Q000235:genetics*; D008297:Male; D035683:MicroRNAs / Q000097:blood* / Q000652:urine; D008875:Middle Aged; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D055815:Young Adult
chemical_list: C505362:MIRN16 microRNA, human; C524940:MIRN21 microRNA, human; C541874:MIRN92 microRNA, human; D035683:MicroRNAs
keywords: Circulating miRNA; Endogenous normalizer; Essential hypertension; Plasma; qRT-PCR