Please use this identifier to cite or link to this item: https://biore.bio.bg.ac.rs/handle/123456789/4022
Title: Hypoxanthine Guanine Phosphoribosyl Transferase Is the Most Stable Reference Gene for Gene Expression Analysis by Quantitative PCR in Peripheral Blood Mononuclear Cells from Women with the Polycystic Ovary Syndrome
Authors: Milutinović, Danijela Vojnović
Macut, Djuro
Antić, Ivana Božić
Macut, Jelica Bjekić
Nikolić, Marina
Matić, Gordana 
Nestorov, Jelena
Issue Date: 2014
Journal: Journal of Medical Biochemistry
Series/Report no.: 33;1-8
Abstract: 
Background: Polycystic ovary syndrome (PCOS) is a frequent
endocrine disorder that affects women in reproductive
age. As the syndrome is strongly associated with obesity,
it is of interest to examine gene expression differences
that accompany its development and associated metabolic
disturbances. Real-time RT PCR is a standard method for
studying changes in gene expression. However, to obtain
accurate and reliable results, validation of reference genes
is obligatory. The aim of this study was to identify suitable
reference for normalization of gene expression in peripheral
blood mononuclear cells (PBMCs) from obese and normal-
weight women with PCOS.
Methods: The expression stability of four potential reference
genes: hypoxanthine guanine phosphoribosyl transferase
1 (HPRT), b-actin (BA), b2-microglobulin (B2M) and
glyceraldehyde-3-phosphate dehydrogenase (GAPDH),
was assessed in PBMCs from healthy women, and from
normal-weight and obese women with PCOS. The variability in
expression of potential reference genes was analyzed
by TaqMan real-time RT PCR method, using GeNorm and
NormFinder software packages.
Results: Direct comparison of cycle threshold (Ct) values
showed inter-individual variations for all validated genes,
the Ct values of HPRT being less variable than those of BA,
GAPDH and B2M. Both software packages pointed to
HPRT as the most steadily expressed gene in PBMCs of
women with PCOS and healthy controls.
Conclusions: Cross-validation of expression stability of four
potential reference genes identified HPRT as the most stable
reference suitable for further investigations of gene
expression in PBMCs from women with PCOS.
URI: https://biore.bio.bg.ac.rs/handle/123456789/4022
ISSN: 1452-8266
DOI: 10.2478/jomb-2014-0029
Appears in Collections:Journal Article

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