人工周期下宫颈脱落细胞基因表达特征与子宫内膜容受性的关联  

Association between gene expression profile of cervical exfoliated cells and endometrial receptivity in artificial cycles

作　　者：杨童焜 彭红梅[2] 杨怡卓[2] 高原[3] 穆莎[2] 董明理 刘佳林 YANG Tongkun;PENG Hongmei;YANG Yizhuo;GAO Yuan;MU Sha;DONG Mingli;LIU Jialin(PLA Medical School,Beijing;Department of Obstetrics and Gynecology,First Medical Center,Chinese PLA General Hospital,Beijing;Department of Obstetrics and Gynecology,Seventh Medical Center,Chinese PLA General Hospital,Beijing,China)

机构地区：[1]解放军医学院,北京 [2]解放军总医院第一医学中心妇产科,北京 [3]解放军总医院第七医学中心妇产科,北京

出　　处：《陆军军医大学学报》2025年第6期571-580,共10页Journal of Army Medical University

基　　金：国家重点研发计划资助(2022YFC2705305)。

摘　　要：目的探讨人工周期条件下宫颈脱落细胞基因表达特征与子宫内膜容受性的潜在关联,以筛选与子宫内膜容受期相关的特异性生物标志物。方法2024年2-10月在解放军总医院第一医学中心生殖中心就诊的准备进行冻融胚胎移植(frozen embryo transfer,FET)的患者中,选取19名患者,根据人工周期冻融胚胎移植方案,在其孕酮使用后第4天(P+4),收集子宫内膜组织验证其子宫内膜容受状态,并在其孕酮使用后第4天(P+4)和第6天(P+6)收集宫颈脱落细胞,利用转录组测序(RNA sequencing,RNA-Seq)检测基因表达情况。将错误发现率阈值(false discovery rate,FDR)小于0.05,Log_(2)差异倍数(fold change,FC)的绝对值大于1作为筛选标准获取差异表达基因(differentially expressed genes,DEGs)并进行生物信息学分析。使用R软件(4.4.1版本)对DEGs进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析,以及蛋白质-蛋白质互作网络(protein-protein interaction,PPI)的构建,再利用Cytoscape软件找寻候选基因。利用定量反转录PCR(quantitative reverse transcription PCR,RT-qPCR)在体内外水平对目标候选基因进行验证。结果子宫内膜容受性测试显示,19例子宫内膜样本在P+6时均达到容受期状态。RNA测序检测分析后,共筛选出3458个DEGs;生信分析结果显示,上调表达的基因与免疫应答、细胞分化通路相关,下调表达的基因与脂质代谢、细胞增殖等相关。利用PPI网络中最大团中心性(maximal clique centrality,MCC)算法选取前20的基因,其中IFIT2、OASL、MX1、RSAD2、IFIT1、IFIT36个基因排并列第一,且这6个基因均属于干扰素刺激基因(interferon-stimulated genes,ISGs)。经实时荧光定量逆转录聚合酶链反应验证,相较于P+6阶段的宫颈脱落细胞,P+4阶段的上述6个基因呈现出更高的表达水平(P<0.05)。结论容受期宫颈脱落细胞的免疫和细胞骨架重塑�Objective To investigate the gene expression profile of cervical exfoliated cells from woman treated by artificial cycle,and their potential association with endometrial receptivity in order to screen specific biomarkers closely related to the receptivity.Methods A total of 19 female patients were enrolled from those preparing for frozen embryo transfer(FET)at the Reproductive Center of First Medical Center of Chinese PLA General Hospital from February 2024 to October 2024.Under the artificial cycle frozen embryo transfer protocol,the endometrial tissues were collected on the 4th day after progesterone administration(P+4)to verify their endometrial receptivity status.Additionally,cervical exfoliated cells were collected on the 4th day(P+4)and the 6th day(P+6)after progesterone administration.RNA sequencing(RNA-Seq)was used to detect gene expression profiles.Differentially expressed genes(DEGs)were identified using the criteria of|log_(2)fold change|>1 and a false discovery rate(FDR)<0.05,followed by bioinformatics analysis.The proteinprotein interaction(PPI)network of DEGs was constructed using R software(4.4.1)and analyzed with gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)analyses.The candidate genes were identified based on the PPI network using Cytoscape software.Quantitative reverse transcription PCR(RT-qPCR)was employed to validate the target candidate genes both in vitro and in vivo.Results The rsERT confirmed that all 19 women were in state of endometrial receptivity at P+6.RNA-Seq identified 3458 DEGs in cervical exfoliated cells between P+4 and P+6.The up-regulated DEGs were mainly enriched in the pathways associated with immune response and cell differentiation,and the down-regulated ones were mainly enriched in the pathways associated with lipid metabolism and cell proliferation.Using maximal clique Centrality(MCC)algorithm in the PPI network,the top 20 genes were selected.Among them,6 genes,such as IFIT2,OASL,MX1,RSAD2,IFIT1 and IFIT3,tied for the first place,and the 6 genes all b

关 键 词：子宫颈脱落细胞 子宫内膜容受性 转录组测序 生物标记物 

分 类 号：R322.65[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学] R711.6