通过 RNA-Seq 检测到人类子宫内膜从预接受状态到接受状态转变过程中的转录组变化。

Transcriptomic changes during the pre-receptive to receptive transition in human endometrium detected by RNA-Seq.

机构信息

Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics (S.G.H., Y.W., H.J.X., X.W.J., Y.Q.H., Q.L.Z., Z.J.C., Y.S.), Center for Reproductive Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200135, China; Shanghai Key Laboratory for Molecular Andrology (S.G.H., G.X.Y.), State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.

出版信息

J Clin Endocrinol Metab. 2014 Dec;99(12):E2744-53. doi: 10.1210/jc.2014-2155.

DOI:10.1210/jc.2014-2155

PMID:25243572

Abstract

CONTEXT

Identifying novel molecular markers for assessing endometrial receptivity is necessary for understanding human implantation and may help in improving the clinical outcome of in vitro fertilization.

OBJECTIVE

We aimed to compare the gene expression profiles of the pre-receptive vs receptive phases of the natural cycle in human endometrial biopsies.

DESIGN

The design of this study was detecting the global gene expression profile of human endometrial receptivity by RNA-Seq.

SETTING

This study was conducted at a university reproductive center.

PARTICIPANTS

Twelve women with normal menstrual cycles participated in the study.

INTERVENTION

Study interventions included endometrial biopsies.

MAIN OUTCOME MEASURES

The endometrial transcriptomes were determined by RNA-Seq, and the expression of selected differentially expressed genes (DEGs) was validated by quantitative RT-PCR.

RESULTS

A total of 2372 DEGs were identified by RNA-Seq. Of these genes, 1099 were up-regulated at LH+7 versus LH+2, whereas 1273 were down-regulated. Nineteen selected genes were confirmed by quantitative RT-PCR. We first demonstrated that metallothionein (MT) family members, MT1E, F, G, H, M, X, and 2A, and four novel transcripts, HAP1, ZCCHC12, MRAP2, and OVGP1, which were not previously linked to endometrial physiology, showed significant expression changes during implantation. Mineral absorption was the most enriched pathway for up-regulated genes, and cell cycle was enriched for down-regulated genes. Gene co-expression network analysis identified five core regulatory factors (GLI2, CDC25A, TLR9, MT1G, and SLC5A1) that are involved in endometrial receptivity during implantation. Examination of the promoter regions of the DEGs identified AP2 and SP1 binding sites, suggesting a potential regulatory role in endometrial gene expression for these two transcription factors.

CONCLUSIONS

This study provides the first RNA-Seq-based transcriptome comparison of pre-receptive and receptive human endometria. Many novel candidate genes, which have not been previously studied in human endometrium, may have functional significance during implantation and serve as molecular markers for endometrial receptivity.

摘要

背景

识别评估子宫内膜容受性的新分子标志物对于理解人类着床非常必要，并且可能有助于改善体外受精的临床结局。

目的

我们旨在比较自然周期中子宫内膜接受期与预接受期的基因表达谱。

设计

本研究通过 RNA-Seq 检测人类子宫内膜容受性的全基因表达谱。

设置

本研究在一所大学生殖中心进行。

参与者

12 名月经周期正常的妇女参与了这项研究。

干预措施

研究干预包括子宫内膜活检。

主要观察指标

通过 RNA-Seq 确定子宫内膜转录组，通过定量 RT-PCR 验证所选差异表达基因（DEG）的表达。

结果

RNA-Seq 共鉴定出 2372 个 DEG。其中，LH+7 与 LH+2 相比，有 1099 个基因上调，1273 个基因下调。通过定量 RT-PCR 验证了 19 个选定基因。我们首次证明，金属硫蛋白（MT）家族成员 MT1E、F、G、H、M、X 和 2A，以及四个新的转录本 HAP1、ZCCHC12、MRAP2 和 OVGP1，这些基因以前与子宫内膜生理无关，在着床过程中表达明显变化。上调基因中最富集的途径是矿物质吸收，下调基因中富集的途径是细胞周期。基因共表达网络分析确定了五个参与着床过程中子宫内膜容受性的核心调节因子（GLI2、CDC25A、TLR9、MT1G 和 SLC5A1）。对 DEG 启动子区域的检查发现了 AP2 和 SP1 结合位点，这表明这两个转录因子在子宫内膜基因表达中可能具有潜在的调节作用。