单细胞RNA测序解析子宫内膜异位症血管生成机制

doi:10.12280/gjfckx.20240982

摘要/Abstract

摘要：

子宫内膜异位症（endometriosis，EMs）作为一种发病机制尚不明确的疾病，亟需借助新兴技术深入阐明其发病过程。将单细胞RNA测序（single-cell RNA sequencing，scRNA-seq）技术应用在EMs研究中，可以从单个细胞信息出发，获取EMs相关的细胞分化、发展、演化及细胞间相互作用等完备信息，帮助对EMs发病机制形成更全面的认知。综述兴起的scRNA-seq技术以及近年来通过scRNA-seq获得的与EMs血管生成领域相关的研究进展。从细胞功能紊乱、免疫失衡等多个影响血管生成的角度出发，探讨EMs相关细胞与血管生成、免疫微环境改变、激素反应性、纤维化等疾病生物学机制之间的关系，以期为进一步探究EMs中血管生成发生机制以及寻找新的治疗靶点提供新思路。

关键词: 序列分析，RNA, 高通量核苷酸序列分析, 子宫内膜异位症, 异质性, 单细胞RNA测序, 血管生成

Abstract:

Endometriosis (EMs), as a disease with an unclear etiology, urgent to clarify its pathogenesis with the help of emerging new technologies. The application of single-cell RNA sequencing (scRNA-seq) technology in the research of EMs can obtain complete information about the differentiation, development, evolution, and intercellular interactions from the perspective of individual cells, to help to form a more comprehensive understanding of the pathogenic mechanism of EMs. This article reviews the emerging scRNA-seq technology and the research progress related to angiogenesis in EMs obtained through scRNA-seq in recent years. From multiple perspectives that affect angiogenesis, such as cellular dysfunction and immune imbalance, this article details the relationship between the associated cells of EMs and disease biological mechanisms such as angiogenesis, immune microenvironment changes, hormone responsiveness, and fibrosis, provides new ideas for exploring the pathogenesis of angiogenesis in EMs and finds new therapeutic targets.

Key words: Sequence analysis, RNA, High-throughput nucleotide sequencing, Endometriosis, Heteroplasmy, Single-cell RNA sequencing, Angiogenesis

曹秀蓉, 周文柏, 范香, 王逸斐, 朱鹏峰. 单细胞RNA测序解析子宫内膜异位症血管生成机制[J]. 国际妇产科学杂志, 2025, 52(2): 199-205.

CAO Xiu-rong, ZHOU Wen-bai, FAN Xiang, WANG Yi-fei, ZHU Peng-feng. Single-Cell RNA Sequencing Analysis of the Angiogenesis Mechanism in Endometriosis[J]. Journal of International Obstetrics and Gynecology, 2025, 52(2): 199-205.

图/表 2

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方法	实验目的	实验流程	数据分析
bulk RNA-seq	检测单个标本中的基因表达情况；比较多个标本间差异表达基因	从整个组织或全部细胞中提取RNA，逆转录，建库测序	评估转录本，基因的表达；差异基因表达分析；可变剪切
scRNA-seq	检测单个细胞中的基因表达情况；比较不同状态，细胞间的表达差异	解离单个细胞加标签，核苷酸序列扩增，建库测序	降维聚类；鉴定细胞亚群；差异基因表达；受体配体互作；拟时序，分化轨迹

方法	实验目的	实验流程	数据分析
bulk RNA-seq	检测单个标本中的基因表达情况；比较多个标本间差异表达基因	从整个组织或全部细胞中提取RNA，逆转录，建库测序	评估转录本，基因的表达；差异基因表达分析；可变剪切
scRNA-seq	检测单个细胞中的基因表达情况；比较不同状态，细胞间的表达差异	解离单个细胞加标签，核苷酸序列扩增，建库测序	降维聚类；鉴定细胞亚群；差异基因表达；受体配体互作；拟时序，分化轨迹

单细胞测序方法	测序原理	区域	UMI	扩增方法	捕获法	优点	缺点
Smart-seq	用含有PCR识别序列的引物和M-MLV RT	全长	×	PCR	流式分选或显微操作	高转录本覆盖，可实现SNP和可变剪切分析	不能分析非poly(A)的RNA；reads不带有mRNA链特异性
Drop-seq	基于微液滴技术，每个cDNA都用一个细胞特异性barcode和UMI标记	3′端	√	PCR	液滴	建库流程快，成本低	需要微流控平台；细胞捕获率较低
CEL-seq	通过逆转录和体外转录整合条形码和UMI；线性扩增	3′端	√	IVT	微流控	错误率比较低	强烈的3′偏好；高丰度转录本被优先扩增；总RNA需求多
10x Genomics	类似于Drop-seq，利用微流体“双十字”交叉系统分选，通过barcode磁珠-单细胞-油滴形成GEM	3′端	√	PCR	液滴	建库流程快，成本低，细胞可适性、捕获效率、通量高	非全长序列；样本要求高（80%活细胞数）
SCRB-seq	使用类似于Smart-seq的PCR扩增，同时集成barcode和UMI	3′端	√	PCR	流式分选	成本低，转录本精确量化，针对体积小、处理步骤少而优化	操作要求高，不能产生全长cDNA，3′偏好类似于CEL-seq

单细胞测序方法	测序原理	区域	UMI	扩增方法	捕获法	优点	缺点
Smart-seq	用含有PCR识别序列的引物和M-MLV RT	全长	×	PCR	流式分选或显微操作	高转录本覆盖，可实现SNP和可变剪切分析	不能分析非poly(A)的RNA；reads不带有mRNA链特异性
Drop-seq	基于微液滴技术，每个cDNA都用一个细胞特异性barcode和UMI标记	3′端	√	PCR	液滴	建库流程快，成本低	需要微流控平台；细胞捕获率较低
CEL-seq	通过逆转录和体外转录整合条形码和UMI；线性扩增	3′端	√	IVT	微流控	错误率比较低	强烈的3′偏好；高丰度转录本被优先扩增；总RNA需求多
10x Genomics	类似于Drop-seq，利用微流体“双十字”交叉系统分选，通过barcode磁珠-单细胞-油滴形成GEM	3′端	√	PCR	液滴	建库流程快，成本低，细胞可适性、捕获效率、通量高	非全长序列；样本要求高（80%活细胞数）
SCRB-seq	使用类似于Smart-seq的PCR扩增，同时集成barcode和UMI	3′端	√	PCR	流式分选	成本低，转录本精确量化，针对体积小、处理步骤少而优化	操作要求高，不能产生全长cDNA，3′偏好类似于CEL-seq