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

YANG Tongkun; PENG Hongmei; YANG Yizhuo; GAO Yuan; MU Sha; DONG Mingli; LIU Jialin

doi:10.16016/j.2097-0927.202412145

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

Clinical Medicine | 更新时间：2025-03-28

- Association between gene expression profile of cervical exfoliated cells and endometrial receptivity in artificial cycles
- Journal of Army Medical University Vol. 47, Issue 6, Pages: 571-580(2025)
- 作者机构：
  
  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
- 作者简介：
  
  PENG Hongmei， E-mail： phmeizys@hotmail.com
- 基金信息：
  
  the National Key Research and Development Program of China(2022YFC2705305)
- DOI：10.16016/j.2097-0927.202412145
  CLC： R322.65;R394.2;R711.6
- Received：30 December 2024，
  
  Revised：21 February 2025，
  
  Published：30 March 2025
- 稿件说明：
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杨童焜，彭红梅，杨怡卓，等.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［J］.陆军军医大学学报，2025，47（06）：571-580.DOI：10.16016/j.2097-0927.202412145.

YANG T K，PENG H M，YANG Y H，et al.Association between gene expression profile of cervical exfoliated cells and endometrial receptivity in artificial cycles［J］.J Army Med Univ，2025，47（06）：571-580.DOI：10.16016/j.2097-0927.202412145.
杨童焜，彭红梅，杨怡卓，等.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［J］.陆军军医大学学报，2025，47（06）：571-580.DOI：10.16016/j.2097-0927.202412145. DOI：

YANG T K，PENG H M，YANG Y H，et al.Association between gene expression profile of cervical exfoliated cells and endometrial receptivity in artificial cycles［J］.J Army Med Univ，2025，47（06）：571-580.DOI：10.16016/j.2097-0927.202412145. DOI：

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

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3 458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、IFIT3

6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（

0.05）。

结论

容受期宫颈脱落细胞的免疫和细胞骨架重塑相关基因表达发生变化，ISGs表达的降低可能作为子宫内膜容受性的潜在生物标志物。

Abstract

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 express

ion profiles. Differentially expressed genes (DEGs) were identified using the criteria of |log

fold change|

1 and a false discovery rate (FDR)

0.05

followed by bioinformatics analysis. The protein-protein 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 3 458 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 belong to interferon-stimulated genes (ISGs). qRT-PCR indicated that the above 6 genes showed significantly higher expression levels in the cervical exfoliated cells at the P+4 stage than the cells at the P+6 stage (

0.05). Conclusion‍　‍There are changes in the expression levels of the genes related to immunity and cytoskeleton remodeling in cervical exfoliated cells during the endometrial receptivity phase. The decrease in the expression of ISGs may serve as a potential biomarker for endometrial receptivity.

关键词

Keywords

references

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