Analysis of Serum microRNA Profile by Solexa Sequencing in Women With Endometriosis
Analysis of Serum microRNA Profile by Solexa Sequencing in Women With Endometriosis
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DOI：
10.1177/1933719116641761
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发表时间：
2016-10-01
影响因子：
2.9
通讯作者：
Xia, Xiaomeng
中科院分区：
文献类型：
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作者：
Wang, Lei;Huang, Wei;Xia, Xiaomeng
目的：应用基因芯片技术研究血清微RNA作为生物标志物在子宫内膜异位症无创性诊断中的潜在作用。然而，微阵列分析在结果准确性和重复性方面表现不佳，不适合用于探索新的靶点。本研究采用Solexa测序法对子宫内膜异位症患者的血清miRNAs谱进行了进一步分析，为将血清miRNAs作为生物标志物应用于子宫内膜异位症的诊断提供了更多的证据。材料和方法：收集30例轻、轻度子宫内膜异位症患者和20例非子宫内膜异位症患者的血清标本。用Solexa测序法检测血清miRNAs的表达，并用实时定量聚合酶链式反应(QPCR)进行验证。结果：Solexa测序结果显示，子宫内膜异位症患者和对照组血清中所有小RNA的清洁率为93.63%。通过深度测序发现，共有108个miRNAs在子宫内膜异位症患者血清中与对照组相比有差异表达。其中，98个miRNAs显著下调，10个miRNAs显著上调。在98个miRNAs中只有21个显著下调，未见文献报道显著上调的miRNAs，这可能是由于样品和分析方法的差异所致。因此，SolexA测序结果在其他样本中得到了qPCR的验证。一些miRNAs被认为是有希望的子宫内膜异位症的诊断标志物。基因本体论和京都基因和基因组百科全书对靶基因的功能注释表明，大多数差异miRNAs可能与子宫内膜异位症有关。结论：循环中的miRNAs可作为早期诊断微轻度子宫内膜异位症的检测生物标志物。
Objective(s): The potential roles of serum microRNAs (miRNAs), as biomarkers, in noninvasive diagnosis of endometriosis have been reported by microarray analysis. However, microarray analysis cannot perform well in outcome accuracy and repeatability and is not suitable to be used for exploring new targets. Here, Solexa sequencing, a wide and precise method, was adopted to further analyze the serum miRNAs profile in endometriosis, which may offer more evidence to apply serum miRNAs as biomarkers in diagnosis of endometriosis.Materials and Methods: Serum samples were collected from 30 patients with minimal-mild endometriosis and 20 women without endometriosis as control. Expression of serum miRNAs was measured by Solexa sequencing and validated by quantitative real-time polymerase chain reaction (qPCR).Results: Solexa sequencing showed 93.63% clean readouts for all small RNAs in the serum of patients with endometriosis and controls. A total of 108 miRNAs were found to be differentially expressed in the serum of patients with endometriosis by deep sequencing, compared to controls. Among them, 98 miRNAs were significantly downregulated, while 10 miRNAs were significantly upregulated. Only 21 of 98 significantly downregulated miRNAs, and none of significantly upregulated miRNAs were reported in published literatures, which may be due to the differences in samples and analytical methods. The Solexa sequencing results were consequently validated by qPCR in additional samples. Some miRNAs were identified to be promising diagnostic markers of endometriosis. The functional annotation of target genes revealed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated that a majority of differential miRNAs might be involved in endometriosis.Conclusion: Circulating miRNAs may be useful as detection biomarkers for the early diagnosis of minimal-mild endometriosis.