NHC Key Laboratory of Carcinogenesis and Hunan Key Laboratory of Cancer Metabolism, Hunan Cancer Hospital and the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, China.
Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute, Central South University, Changsha, China.
J Exp Clin Cancer Res. 2021 May 11;40(1):163. doi: 10.1186/s13046-021-01955-1.
Single-cell RNA sequencing (scRNA-seq) is a tool for studying gene expression at the single-cell level that has been widely used due to its unprecedented high resolution. In the present review, we outline the preparation process and sequencing platforms for the scRNA-seq analysis of solid tumor specimens and discuss the main steps and methods used during data analysis, including quality control, batch-effect correction, normalization, cell cycle phase assignment, clustering, cell trajectory and pseudo-time reconstruction, differential expression analysis and gene set enrichment analysis, as well as gene regulatory network inference. Traditional bulk RNA sequencing does not address the heterogeneity within and between tumors, and since the development of the first scRNA-seq technique, this approach has been widely used in cancer research to better understand cancer cell biology and pathogenetic mechanisms. ScRNA-seq has been of great significance for the development of targeted therapy and immunotherapy. In the second part of this review, we focus on the application of scRNA-seq in solid tumors, and summarize the findings and achievements in tumor research afforded by its use. ScRNA-seq holds promise for improving our understanding of the molecular characteristics of cancer, and potentially contributing to improved diagnosis, prognosis, and therapeutics.
单细胞 RNA 测序（scRNA-seq）是一种用于研究单细胞水平基因表达的工具，由于其前所未有的高分辨率，因此得到了广泛应用。在本综述中，我们概述了用于固体肿瘤标本 scRNA-seq 分析的准备过程和测序平台，并讨论了数据分析过程中的主要步骤和方法，包括质量控制、批次效应校正、标准化、细胞周期相位分配、聚类、细胞轨迹和伪时间重建、差异表达分析和基因集富集分析，以及基因调控网络推断。传统的批量 RNA 测序不能解决肿瘤内和肿瘤间的异质性，自第一个 scRNA-seq 技术发展以来，这种方法已被广泛应用于癌症研究，以更好地理解癌细胞生物学和发病机制。scRNA-seq 对靶向治疗和免疫治疗的发展具有重要意义。在本综述的第二部分，我们重点介绍了 scRNA-seq 在实体肿瘤中的应用，并总结了其在肿瘤研究中发现和取得的成果。scRNA-seq 有望提高我们对癌症分子特征的理解，并有可能促进诊断、预后和治疗的改善。
