Integrated Transcriptomic Analysis of the miRNA-mRNA Interaction Network in Thin Endometrium [RNA-Seq]

Although the thin endometrium (TE) has been widely recognized as a critical factor in implantation failure, the contribution of miRNA-mRNA regulatory network to the development of disease aetiology remains to be further elucidated. This study performed an integrative analysis of the miRNA-mRNA expression profiles in the thin and adjacent normal endometrium of 8 patients with intrauterine adhesion to construct the transcriptomic regulatory networks. A total of 1093 differentially expressed genes (DEGs) and 72 differentially expressed miRNAs (DEMs) were identified in the thin adhesive endometrium of the TE group compared with the control adjacent normal endometrial cells. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed that DEGs and the target genes of DEM were significantly enriched in angiogenesis, cell growth regulation and Wnt signalling pathway. Multiple hub genes (CAV1, MET, MAL2, has-mir-138, ARHGAP6, CLIC4, RRAS, AGFG1, has-mir-200, and has-mir-429) were identified by constructing the miRNA-mRNA regulatory networks. Furthermore, a miRNA-mRNA-pathway-function analysis was conducted, and the hub genes were enriched in the FoxO signalling pathway, cell growth regulation, inflammatory response regulation and regulation of autophagy pathways. Our study is the first to perform integrated mRNA-seq and miRNA-seq analyses in the thin adhesive endometrium and the control adjacent normal endometrial cells. This study provides new insights into the molecular mechanisms underlying the formation of thin endometrium. thin adhesive endometrium and the control adjacent normal endometrial cells of 8 patients with intrauterine adhesion.The thin endometrium RNA and adjacent normal endometrial RNA respective pooled equally, and reverse transcribed into cDNAs using the QuantiTect Reverse Transcription Kit.

尽管薄型子宫内膜（thin endometrium, TE）已被广泛认为是导致着床失败的关键危险因素，但miRNA-mRNA调控网络在该疾病病因学发生发展中的作用仍有待进一步阐明。本研究对8例宫腔粘连患者的薄型粘连子宫内膜及其邻近正常子宫内膜组织的miRNA-mRNA表达谱进行整合分析，以构建转录组调控网络。与对照组邻近正常子宫内膜细胞相比，TE组的薄型粘连子宫内膜中共鉴定出1093个差异表达基因（differentially expressed genes, DEGs）和72个差异表达miRNA（differentially expressed miRNAs, DEMs）。基因本体（Gene Ontology, GO）及京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路富集分析结果显示，DEGs以及DEMs的靶基因显著富集于血管生成、细胞生长调控以及Wnt信号通路。通过构建miRNA-mRNA调控网络，本研究鉴定出多个核心基因，包括CAV1、MET、MAL2、has-mir-138、ARHGAP6、CLIC4、RRAS、AGFG1、has-mir-200以及has-mir-429。此外，本研究还开展了miRNA-mRNA-通路-功能分析，结果显示核心基因富集于FoxO信号通路、细胞生长调控、炎症反应调控以及自噬调控通路。本研究首次在宫腔粘连患者的薄型粘连子宫内膜及邻近正常子宫内膜细胞中开展整合mRNA测序与miRNA测序分析，为阐明薄型子宫内膜形成的分子机制提供了新的视角。将8例宫腔粘连患者的薄型粘连子宫内膜RNA与邻近正常子宫内膜RNA分别等量混合，采用QuantiTect反转录试剂盒将其反转录为cDNA。