A high-throughput open-array qPCR gene panel to identify housekeeping genes suitable for myometrium and leiomyoma expression analysis.
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.
Instituto de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Campus de Anchieta, Avda. Astrofísico Francisco Sánchez s/n, La Laguna, Tenerife 38071, Spain. Electronic address: tacosalm@ull.edu.es.
Teresa A
TA
Almeida
Instituto de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Campus de Anchieta, Avda. Astrofísico Francisco Sánchez s/n, La Laguna, Tenerife 38071, Spain. Electronic address: maquispe@ull.es.
Antonieta
A
Quispe-Ricalde
Hospital Quirón, Poeta Rodríguez Herrera 1, Santa Cruz de Tenerife, Tenerife 38006, Spain. Electronic address: fmontesdeoca@ginefem.com.
Francisco
F
Montes de Oca
Instituto de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Campus de Anchieta, Avda. Astrofísico Francisco Sánchez s/n, La Laguna, Tenerife 38071, Spain. Electronic address: pforonda@ull.es.
Pilar
P
Foronda
Instituto de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, Campus de Anchieta, Avda. Astrofísico Francisco Sánchez s/n, La Laguna, Tenerife 38071, Spain. Electronic address: mnhdez@ull.edu.es.
Mariano M
MM
Hernández
D000328:Adult; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007044:Hysterectomy; D007889:Leiomyoma / Q000235:genetics* / Q000378:metabolism / Q000601:surgery; D008875:Middle Aged; D009215:Myometrium / Q000378:metabolism / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D014594:Uterine Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000601:surgery
Housekeeping; Leiomyoma; Myometrium; Open-array panel; qPCR