MicroRNAs: new candidates for the regulation of the human cumulus-oocyte complex
MicroRNAs: new candidates for the regulation of the human cumulus-oocyte complex
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DOI：
10.1093/humrep/det321
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发表时间：
2013-11-01
影响因子：
6.1
通讯作者：
Hamamah, S.
中科院分区：
文献类型：
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作者：
Assou, S.;Al-edani, T.;Hamamah, S.
人类卵丘卵母细胞复合体（COCs）中microRNAs（miRNAs）的表达模式是什么？几种miRNA在卵丘细胞（CC）或卵母细胞中富集，并且被预测为靶向参与COC的生物学功能的基因。然而，关于转录后调节因子（如miRNA）的数据非常有限。这是第一项使用深度测序方法对人类卵母细胞和CC中的小RNA（包括miRNA）进行鉴定和定量的研究。MII卵母细胞和CC收集自接受IVF的女性。使用Illumina/深度测序技术，我们分析了合并的MII卵母细胞（n 24）和CC样本（n 20）的小RNA组。使用计算机预测算法鉴定CC和MII卵母细胞miRNA的mRNA靶标。使用寡核苷酸芯片，全基因组基因表达进行了研究，在卵母细胞（10池19 - 3卵母细胞/每个）和10个单独的CC样品。使用TaqMan miRNA测定来确认MII卵母细胞（3个池，每个8 3个卵母细胞）和CC样品（3个池，每个7 3个CC）的独立池中的测序结果。在人类CC的原代培养物中评估了一种miRNA MIR 23 a的功能作用，小RNA的深度测序产生了超过100万个原始读数。通过将具有单个位置的读段映射到人类基因组，鉴定了在MII卵母细胞（MIR 184、MIR 100和MIR 10A）或CC（MIR 29 a、MIR 30 d、MIR 21、MIR 93、MIR 320 a、MIR 125 a和LET 7家族）中丰富的已知miRNA。预测的卵母细胞miRNAs靶基因与转录和细胞周期调控相关，而CC miRNAs靶基因与细胞外基质和细胞凋亡相关。比较预测的miRNA靶基因和mRNA微阵列数据，得到了224个在MII卵母细胞和CC中差异表达的靶基因，包括PTGS 2，CTGF和BMPR 1B，它们对卵丘卵母细胞通讯很重要。利用原代培养的CC进行的功能分析显示，当MIR 23 a过表达时，BCL 2和CYP 19 A1 mRNA水平降低。此外，材料的来源是MII卵母细胞未能受精。目前的研究结果表明，miRNA可能在卵母细胞和CC crosstalk的调节中发挥作用。这项工作得到了Ferring Pharmaceuticals的部分资助。本研究的作者没有利益冲突需要报告。不适用。
What is the expression pattern of microRNAs (miRNAs) in human cumulusoocyte complexes (COCs)?Several miRNAs are enriched in cumulus cells (CCs) or oocytes, and are predicted to target genes involved in biological functions of the COC.The transcriptional profiles of human MII oocytes and the surrounding CCs are known. However, very limited data are available about post-transcriptional regulators, such as miRNAs. This is the first study focussing on the identification and quantification of small RNAs, including miRNAs, in human oocytes and CCs using a deep-sequencing approach.MII oocytes and CCs were collected from women who underwent IVF.Using the Illumina/deep-sequencing technology, we analyzed the small RNAome of pooled MII oocytes (n 24) and CC samples (n 20). The mRNA targets of CC and MII oocyte miRNAs were identified using in silico prediction algorithms. Using oligonucleotide microarrays, genome-wide gene expression was studied in oocytes (10 pools of 19 3 oocytes/each) and 10 individual CC samples. TaqMan miRNA assays were used to confirm the sequencing results in independent pools of MII oocytes (3 pools of 8 3 oocytes/each) and CC samples (3 pools of 7 3 CCs/each). The functional role of one miRNA, MIR23a, was assessed in primary cultures of human CCs.Deep sequencing of small RNAs yielded more than 1 million raw reads. By mapping reads with a single location to the human genome, known miRNAs that were abundant in MII oocytes (MIR184, MIR100 and MIR10A) or CCs (MIR29a, MIR30d, MIR21, MIR93, MIR320a, MIR125a and the LET7 family) were identified. Predicted target genes of the oocyte miRNAs were associated with the regulation of transcription and cell cycle, whereas genes targeted by CC miRNAs were involved in extracellular matrix and apoptosis. Comparison of the predicted miRNA target genes and mRNA microarray data resulted in a list of 224 target genes that were differentially expressed in MII oocytes and CCs, including PTGS2, CTGF and BMPR1B that are important for cumulusoocyte communication. Functional analysis using primary CC cultures revealed that BCL2 and CYP19A1 mRNA levels were decreased upon MIR23a overexpression.Only known miRNAs were investigated in the present study on COCs. Moreover, the source of the material is MII oocytes that failed to fertilize.The present findings suggest that miRNA could play a role in the regulation of the oocyte and CC crosstalk.This work was partially supported by a grant from Ferring Pharmaceuticals. The authors of the study have no conflict of interest to report.Not applicable.