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

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 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miR-146a-5p的表达水平分别为8.3±10.6、1.9±0.8，对照组分别为1.1±0.6、0.9±0.4，两组分别比较，差异均有统计学意义（P=0.027，P=0.015），与miRNA芯片筛查结果一致。内异症组卵泡期内膜组织中miR-543的表达水平（2.3±1.0）显著高于对照组（1.0±0.4，P=0.008）；但其着床窗口期内膜组织中miR-543的表达水平（1.2±0.6）与对照组（1.5±1.0）比较，差异则无统计学意义（P=0.890）。结论内异症不孕患者着床窗口期内膜组织中存在多个差异表达的miRNA，其中miR-142-5p和miR-146a-5p表达明显上调，可能通过作用于多种子宫内膜容受性标志分子影响胚胎着床；内异症不孕患者着床窗口期内膜组织中miR-543的表达低于卵泡期，提示miR-543可能是影响子宫内膜容受性的原因。 更多相关知识 abstractsObjective 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. Sixty-three differentially expressed mRNA were indentified, among which 58 mRNA such as CADM1, IL-10RA, ITGAL and LPAR5 had significantly enhanced expression. Five mRNA such as HLA-DRB1,3,4,5 and SOHLH2 showed significantly inhibited expression. Thirty-six taget genes were found in consideration of negatively correlated miRNA expression with the genes, miRNA-mRNA co-expression network were constructed. The miR-543 was found at the core of the network. Targetscan and other database predicted that miR-142-5p, miR-146a-5p and miR-543 could act on various types of endometrial receptivity molecular corresponding marker genes such as HOX10, ITGAV, ITGB3, OPN, LIF, ESR, PGR, CDH1 and MMP. (2) RT-PCR test showed that the average levels of expression of miR-142-5p and miR-146a-5p in implantation window phase endometrium in endometriosis group were 8.3 ± 10.6 and 1.9 ± 0.8 respectively;the average levels of that in control group were 1.1±0.6 and 0.9±0.4, respectively. The difference was statistically significant (P=0.027, P=0.015), and was consistent with results from miRNA microarray test. The expression of miR-543 in tissues of follicular phase endometrium in endometriosis group (2.3±1.0) was significantly higher than that in control group (1.0 ± 0.4), and the difference was statistically significant (P=0.008). However, when comparing the expression of miR-543 implantation window phase endometrium in endometriosis group (1.2±0.6) with that in control group (1.5 ± 1.0), the difference was not statistical significant (P=0.890). Conclusions There are multiple differential expressions of miRNA in the implantation window phase endometrium tissues of endometriosis infertility patients, among which miR-142-5p and miR-146a-5p show significantly enhanced expression and may affect embryo implantation by acting on a variety of endometrial receptivity marker molecules. The expression of miR-543 in implantation window phase endometrium of endometriosis infertility patients is lower than that in the follicular phase, forewarned changes in the pattern of cyclic variation of miR-543, and may be the reason for affecting endometrial receptivity. More相关知识 - 浏览0 - 被引26 - 下载0 相似文献 - 中文期刊 - 外文期刊 - 学位论文 - 会议论文