A high-throughput open-array qPCR gene panel to identify housekeeping genes suitable for myometrium and leiomyoma expression analysis

Abstract Objective To evaluate 51 different housekeeping genes for their use as internal standards in myometrial and matched leiomyoma samples in proliferative and secretory phases. Methods RNA from 6 myometrium and matched leiomyoma samples was obtained from pre-menopausal women who underwent hyste...

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Bibliographic Details
Published in:Gynecologic oncology 2014-07, Vol.134 (1), p.138-143
Main Authors: Almeida, Teresa A, Quispe-Ricalde, Antonieta, Montes de Oca, Francisco, Foronda, Pilar, Hernández, Mariano M
Format: Article
Language:English
Subjects:
Adult
Female
Hematology, Oncology and Palliative Medicine
High-Throughput Nucleotide Sequencing - methods
Housekeeping
Humans
Hysterectomy
Leiomyoma
Leiomyoma - genetics
Leiomyoma - metabolism
Leiomyoma - surgery
Middle Aged
Myometrium
Myometrium - metabolism
Myometrium - physiology
Obstetrics and Gynecology
Open-array panel
qPCR
Real-Time Polymerase Chain Reaction - methods
Uterine Neoplasms - genetics
Uterine Neoplasms - metabolism
Uterine Neoplasms - surgery
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Summary:Abstract Objective To evaluate 51 different housekeeping genes for their use as internal standards in myometrial and matched leiomyoma samples in proliferative and secretory phases. Methods RNA from 6 myometrium and matched leiomyoma samples was obtained from pre-menopausal women who underwent hysterectomy. Reverse-transcription and real-time quantitative PCR were achieved using TaqMan high-density open-array human endogenous control panel. Results Expression stability of 51 candidate genes was determined by GeNorm and NormFinder softwares. We identified 10 housekeeping genes, ARF1, MRPL19, FBXW2, PUM1, UBE2D2, EIF2B1, HPRT1, GUSB, ALAS1, and TRIM27, as the best set of normalization genes for comparing relative expression between leiomyoma and myometrium samples in proliferative and secretory phases. Conclusions Adequate reference genes for accurate normalization are essential to compare gene expression between leiomyoma and myometrial samples. Ideal housekeeping genes must have stable expression patterns regardless of the sample type and menstrual cycle phase. In this study, we propose a set of 10 candidate genes with greater expression stability than those housekeeping genes commonly used in leiomyoma and myometrium tissues. Their use will improve the sensitivity and specificity of the gene expression analysis in these tissues.
ISSN:0090-8258
1095-6859
DOI:10.1016/j.ygyno.2014.04.012