Altered Profile of Seminal Plasma MicroRNAs in the Molecular Diagnosis of Male Infertility
Altered Profile of Seminal Plasma MicroRNAs in the Molecular Diagnosis of Male Infertility
复制标题
男性不育分子诊断中精浆 microRNA 谱的改变
DOI：
10.1373/clinchem.2011.169714
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发表时间：
2011-12-01
影响因子：
9.3
通讯作者：
Zhang, Chunni
中科院分区：
文献类型：
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作者：
Wang, Cheng;Yang, Cuihua;Zhang, Chunni
背景：虽然microRNAs(MiRNAs)在精子发生中起着重要作用，但对不育男性精浆miRNAs的研究却知之甚少。我们调查了不育男性精浆miRNAs的谱，以确定在不育患者中发生改变的miRNAs，并评估其诊断价值。方法：采集289例不育男性和168例年龄匹配的正常生育男性精浆样本。首先通过时间历程和冻融循环分析来评估miRNAs的稳定性。使用Solexa测序技术对来自45名非梗阻性无精子症患者、58名弱精子症患者和100名生育对照组的样本中的miRNAs进行了初步筛选。用茎环定量逆转录聚合酶链式反应(RT-qPCR)检测73例非梗阻性无精子症患者、79例弱精子症患者、34例少精子症患者和68例生育对照组精浆中miRNAs的浓度。SolexA测序分析显示，与对照组相比，患者组中有19个miRNAs发生了显著变化。RT-qPCR分析发现，7个miRNAs(miR-34c-5p、miR-122、miR-146b-5p、miR-181a、miR-374b、miR-509-5p和miR-513a-5p)在无精症患者显著减少，在弱精子症患者显著增加。这些miRNA的ROC曲线下面积为0.733~0.921，明显高于常规生化指标(0.510~0.622)。结论：精浆miRNAs的检测为男性不育症的诊断提供了一种新的、无创的方法。(C)2011年美国临床化学协会
BACKGROUND: Although microRNAs (miRNAs) play essential roles in spermatogenesis, little is known about seminal plasma miRNAs in infertile men. We investigated the profile of seminal plasma miRNAs in infertile men to identify miRNAs that are altered in infertility; we then evaluated their diagnostic value.METHODS: Seminal plasma samples were obtained from 289 infertile men and 168 age-matched fertile control individuals. The stability of the miRNAs was first assessed by time-course and freeze-thaw cycle analyses. The Solexa sequencing technology was used for an initial screen of the miRNAs in samples pooled from 45 patients with nonobstructive azoospermia, 58 patients with asthenozoospermia, and 100 fertile controls. A stem-loop quantitative reverse-transcription PCR (RT-qPCR) assay was conducted in the training and verification sets to confirm the concentrations of the altered miRNAs in 73 patients with nonobstructive azoospermia, 79 patients with asthenozoospermia, 34 patients with oligospermia, and 68 fertile controls.RESULTS: The miRNAs in seminal plasma were stable. The Solexa sequencing analysis demonstrated 19 markedly altered miRNAs in the patient groups, compared with the control group. RT-qPCR analysis identified 7 miRNAs (miR-34c-5p, miR-122, miR-146b-5p, miR-181a, miR-374b, miR-509-5p, and miR-513a-5p) as markedly decreased in azoospermia but increased in asthenozoospermia. The area under the ROC curve for these miRNAs ranged from 0.733 to 0.921, markedly higher than for routine biochemical parameters (0.510-0.622). Moreover, the concentrations of some selected miRNAs were also increased in the semen sperm of the asthenozoospermia patients.CONCLUSIONS: The measurement of miRNAs in seminal plasma provides a novel, noninvasive approach for diagnosing male infertility. (C) 2011 American Association for Clinical Chemistry