子宫内膜异位症不孕患者着床窗口期内膜组织中微小RNA的差异表达及其意义  

Differential expression of microRNA in eutopic endometrium tissue during implantation window for patients with endometriosis related infertility

作　　者：王洋[1] 马彩虹[1] 乔杰[1] 

机构地区：[1]北京大学第三医院生殖医学中心,100191

出　　处：《中华妇产科杂志》2016年第6期436-441,共6页Chinese Journal of Obstetrics and Gynecology

基　　金：国家自然科学基金（81170618）

摘　　要：目的：通过检测子宫内膜异位症（内异症）不孕患者着床窗口期子宫内膜组织中差异表达的微小RNA（miRNA），预测影响子宫内膜容受性的相关基因。方法收集2013年5—12月因“不孕症”就诊于北京大学第三医院生殖医学中心的患者，选择其中与内异症相关的不孕患者（17例，包括6例卵泡期内膜、11例着床窗口期内膜）作为内异症组，因输卵管因素不孕的患者（19例，包括7例卵泡期内膜、12例着床窗口期内膜）作为对照组。（1）两组患者各取3例着床窗口期内膜，采用miRNA和mRNA芯片技术筛选，联合数据库预测结果以及miRNA和mRNA的负调控关系，得到与miRNA表达呈负相关的交集靶基因，构建miRNA-mRNA共表达网络，结合生物信息学方法预测子宫内膜容受性相关基因，找到处于核心调控地位的miRNA。（2）采用逆转录（RT）-PCR技术检测两组（内异症组和对照组分别为14例和16例）子宫内膜组织中miR-142-5p、miR-146a-5p和miR-543的表达，验证miRNA芯片技术筛查结果。结果（1）miRNA和mRNA芯片技术筛选结果显示，内异症组着床窗口期内膜组织中，筛选出6个差异表达的miRNA，其中miR-142-5p、miR-146a-5p、miR-1281、miR-940、miR-4634表达明显上调，miR-543表达明显下调；筛选出63个差异表达的基因mRNA，其中CADM1、IL-10RA、ITGAL、LPAR5等58个mRNA表达明显上调，HLA-DRB1、3、4、5和SOHLH2等5个mRNA表达明显下调。联合基因测序结果、数据库预测结果以及miRNA和mRNA的负调控关系，得到与miRNA表达呈负相关的交集靶基因36个，构建miRNA-mRNA共表达网络，发现miR-543处于网络的核心位置；Targetscan等数据库预测miR-142-5p、miR-146a-5p和miR-543可作用于HOX10、ITGAV、ITGB3、OPN、LIF、ESR、PGR、CDH1和MMP等多种子宫内膜容受性标志分子对应的基因。（2）RT-PCR技术检测显示，内异症组着床窗口期内膜组织中miR-142-5p和miObjective To predict the genes that affect endometrial receptivity through the differential expression of microRNA （miRNA） in eutopic endometrial tissues during implantation window in patients with endometriosis infertility. Methods Among infertile patients that received treatments at the Center for Reproductive Medicine,Peking University Third Hospital between May and December 2013, patients with endometriosis infertility were selected as endometriosis group （among the selected 17 cases, there were 6 cases with follicular phase endometrium and 11 cases with implantation window phase endometrium）, and patients with tubal factor infertility were selected as control group （among the selected 19 cases, there were 7 cases with follicular phase endometrium and 12 cases with implantation window phase endometrium）. （1）Implantation window phase endometrium was selected from 3 cases in each group. Using miRNA and mRNA joint gene sequencing and database for forecast results, as well as using the negative regulatory relationship between miRNA and mRNA, the intersection of target gene that negatively correlates with miRNA expression were obtained. The co-expression network of miRNA-mRNA wae constructed. Combined with the genes associated with endometrial receptivity found through bioinformatics method, the miRNA with core regulatory functionwas found. （2） Expand sample size to 14 cases for endometriosis group and 16 cases for control group.Reverse transcription （RT）-PCR technique was utilized to detect the expression of miR-142-5p, miR-146a-5p and miR-543 in endometrial tissues, and verify miRNA microarray results. Results （1） miRNA and mRNA microarray screening results showed that, among the endometrial tissues of patients with endometriosis infertility and with implantation window phase, 6 differentially expressed miRNA were indentified, among which miR-142-5p, miR-146a-5p, miR-1281, miR-940, miR-4634 showed significantly enhanced expression and miR-543 showed significantly inhibited expression.

关 键 词：子宫内膜异位症 不育 女(雌)性 卵泡期 微RNAS RNA 信使 

分 类 号：R711.71[医药卫生—妇产科学] R711.6[医药卫生—临床医学]