title: Identification of miR-23a as a novel microRNA normalizer for relative quantification in human uterine cervical tissues.
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 dereg-ulation 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.
authors.affiliation: Women's Reproductive Health Laboratory of Zhejiang Province.
authors.forename: Yuanming
authors.initials: Y
authors.lastname: Shen
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.forename: Feng
authors.initials: F
authors.lastname: Ye
authors.forename: Fenfen
authors.initials: F
authors.lastname: Wang
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wan
authors.forename: Weiguo
authors.initials: W
authors.lastname: Lu
authors.forename: Xing
authors.initials: X
authors.lastname: Xie
mesh_terms: D002584:Cervix Uteri / Q000378:metabolism* / Q000473:pathology; D055088:Early Detection of Cancer; D005260:Female; D020869:Gene Expression Profiling / Q000592:standards*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism* / Q000592:standards; D046228:Microarray Analysis; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology
chemical_list: C552430:MIRN23a microRNA, human; C554214:MIRN424 microrna, human; D035683:MicroRNAs