多囊卵巢综合征患者分离出的卵丘细胞中长链非编码RNA的异常表达。
Aberrant expression of long noncoding RNAs in cumulus cells isolated from PCOS patients.
作者信息
Huang Xin, Hao Cuifang, Bao Hongchu, Wang Meimei, Dai Huangguan
机构信息
Reproductive Medicine Centre, Affiliated Hospital of Qingdao Medical University, Yuhuangding Hospital of Yantai, 20 Yuhuangding Road East, Yantai, Shandong, 264000, People's Republic of China.
出版信息
J Assist Reprod Genet. 2016 Jan;33(1):111-21. doi: 10.1007/s10815-015-0630-z. Epub 2015 Dec 9.
PURPOSE
To describe the long noncoding RNA (lncRNA) profiles in cumulus cells isolated from polycystic ovary syndrome (PCOS) patients by employing a microarray and in-depth bioinformatics analysis. This information will help us understand the occurrence and development of PCOS.
METHODS
In this study, we used a microarray to describe lncRNA profiles in cumulus cells isolated from ten patients (five PCOS and five normal women). Several differentially expressed lncRNAs were chosen to validate the microarray results by quantitative RT-PCR (qRT-PCR). Then, the differentially expressed lncRNAs were classified into three subgroups (HOX loci lncRNA, enhancer-like lncRNA, and lincRNA) to deduce their potential features. Furthermore, a lncRNA/mRNA co-expression network was constructed by using the Cytoscape software (V2.8.3, http://www.cytoscape.org/ ).
RESULTS
We observed that 623 lncRNAs and 260 messenger RNAs (mRNAs) were significantly up- or down-regulated (≥2-fold change), and these differences could be used to discriminate cumulus cells of PCOS from those of normal patients. Five differentially expressed lncRNAs (XLOC_011402, ENST00000454271, ENST00000433673, ENST00000450294, and ENST00000432431) were selected to validate the microarray results using quantitative RT-PCR (qRT-PCR). The qRT-PCR results were consistent with the microarray data. Further analysis indicated that many differentially expressed lncRNAs were transcribed from chromosome 2 and may act as enhancers to regulate their neighboring protein-coding genes. Forty-three lncRNAs and 29 mRNAs were used to construct the coding-non-coding gene co-expression network. Most pairs positively correlated, and one mRNA correlated with one or more lncRNAs.
CONCLUSIONS
Our study is the first to determine genome-wide lncRNA expression patterns in cumulus cells isolated from PCOS patients by microarray. The results show that clusters of lncRNAs were aberrantly expressed in cumulus cells of PCOS patients compared with those of normal women, which revealed that lncRNAs differentially expressed in PCOS and normal women may contribute to the occurrence of PCOS and affect oocyte development.
目的
通过微阵列和深入的生物信息学分析，描述从多囊卵巢综合征（PCOS）患者分离的卵丘细胞中的长链非编码RNA（lncRNA）谱。这些信息将有助于我们了解PCOS的发生和发展。
方法
在本研究中，我们使用微阵列描述从10名患者（5名PCOS患者和5名正常女性）分离的卵丘细胞中的lncRNA谱。选择几个差异表达的lncRNAs，通过定量逆转录聚合酶链反应（qRT-PCR）验证微阵列结果。然后，将差异表达的lncRNAs分为三个亚组（HOX基因座lncRNA、增强子样lncRNA和长链间质性非编码RNA）以推断其潜在特征。此外，使用Cytoscape软件（V2.8.3，http://www.cytoscape.org/）构建lncRNA/mRNA共表达网络。
结果
我们观察到623个lncRNAs和260个信使RNA（mRNAs）显著上调或下调（≥2倍变化），这些差异可用于区分PCOS患者和正常患者的卵丘细胞。选择5个差异表达的lncRNAs（XLOC_011402、ENST00000454271、ENST00000433673、ENST00000450294和ENST00000432431），使用定量RT-PCR（qRT-PCR）验证微阵列结果。qRT-PCR结果与微阵列数据一致。进一步分析表明，许多差异表达的lncRNAs从2号染色体转录，可能作为增强子调节其邻近的蛋白质编码基因。使用43个lncRNAs和29个mRNAs构建编码-非编码基因共表达网络。大多数对呈正相关，一个mRNA与一个或多个lncRNAs相关。
结论
我们的研究首次通过微阵列确定了从PCOS患者分离的卵丘细胞中的全基因组lncRNA表达模式。结果表明，与正常女性相比，PCOS患者卵丘细胞中lncRNAs簇异常表达，这表明PCOS患者和正常女性中差异表达的lncRNAs可能导致PCOS的发生并影响卵母细胞发育。
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