Identification of miR-23a as a novel microRNA normalizer for relative quantification in human uterine cervical tissues
Experimental & Molecular Medicine
; : 358-366, 2011.
Article
в En
| WPRIM
| ID: wpr-121323
Ответственная библиотека:
WPRO
ABSTRACT
Quantitative real-time RT-PCR (RT-qPCR) is being widely used in microRNA expression research. However, few reports detailed a robust identification and validation strategy for suitable reference genes for normalisation in microRNA RT-qPCR studies. The aim of this study was to identify the most stable reference gene(s) for quantification of microRNA expression analysis in uterine cervical tissues. A microarray was performed on 6 pairs of uterine cervical tissues to identify the candidate reference genes. The stability of candidate reference genes was assessed by RT-qPCR in 23 pairs of uterine cervical tissues. The identified most stable reference genes were further validated in other cohort of 108 clinical uterine cervical samples: (HR-HPV- normal, n = 21; HR-HPV+ normal, n = 19; cervical intraepithelial neoplasia [CIN], n = 47; cancer, n = 21), and the effects of normalizers on the relative quantity of target miR-424 were assessed. In the array experiment, miR-26a, miR-23a, miR-200c, let-7a, and miR-1979 were identified as candidate reference genes for subsequent validation. MiR-23a was identified as the most reliable reference gene followed by miR-191. The use of miR-23a and miR-191 to normalize expression data enabled detection of a significant deregulation of miR-424 between normal, CIN and cancer tissue. Our results suggested that miR-23a and miR-191 are the optimal reference microRNAs that can be used for normalization in profiling studies of cervical tissues; miR-23a is a novel microRNA normalizer.
Key words
Полный текст:
1
База данных:
WPRIM
Основная тема:
Reference Standards
/
Uterine Cervical Dysplasia
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Uterine Cervical Neoplasms
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Cervix Uteri
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Reverse Transcriptase Polymerase Chain Reaction
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Gene Expression Profiling
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MicroRNAs
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Microarray Analysis
/
Early Detection of Cancer
Тип исследования:
Diagnostic_studies
/
Prognostic_studies
/
Screening_studies
Пределы темы:
Female
/
Humans
Язык:
En
Журнал:
Experimental & Molecular Medicine
Год:
2011
Тип:
Article