Cancer Molecular Pathology of School of Medicine, Griffith University, Gold Coast, Queensland, Australia.
Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh.
Methods Mol Biol. 2020;2129:177-191. doi: 10.1007/978-1-0716-0377-2_14.
Cancer stem cells (CSCs) are a small subpopulation of cells associated with cancer initiation, progression, metastasis, therapy resistant, and recurrence. In esophageal squamous cell carcinoma (ESCC), several cell surface and intracellular markers, for example, CD44, ALDH, Pygo2, MAML1, Twist1, Musashi1, side population (SP), CD271, and CD90, have been proposed to identify CSCs. In addition, stem cell markers such as ALDH1, HIWI, Oct3/4, ABCG2, SOX2, SALL4, BMI-1, NANOG, CD133, and podoplanin were associated with pathological stages of cancer, cancer recurrence, prognosis, and therapy resistance of patients with ESCC. Identification and isolation of CSCs could play an important part of improved cancer management regime in ESCC. Furthermore, CSCs may be used as the predictive tool for chemoradiotherapy response in ESCC. Different methods such as in vitro functional assays, cell sorting using various intracellular, and cell surface markers and xenotransplantation techniques are frequently used for the identification and isolation of CSCs in different cancers, including ESCC. However, none of these methods solely can guarantee complete isolation of CSC population. Therefore, a combination of methods is used for reliable detection and isolation of CSCs. Herein, we describe the identification and isolation of CSCs from ESCC cells by cell sorting after Hoechst 33342 staining followed by in vitro functional assays and in vivo mouse xenotransplantation techniques.
癌症干细胞（CSCs）是与癌症起始、进展、转移、治疗耐药和复发相关的一小部分细胞。在食管鳞状细胞癌（ESCC）中，已经提出了几种细胞表面和细胞内标志物，例如 CD44、ALDH、Pygo2、MAML1、Twist1、Musashi1、侧群（SP）、CD271 和 CD90，用于鉴定 CSCs。此外，干细胞标志物，如 ALDH1、HIWI、Oct3/4、ABCG2、SOX2、SALL4、BMI-1、NANOG、CD133 和 podoplanin，与 ESCC 患者的癌症病理分期、癌症复发、预后和治疗耐药性相关。CSCs 的鉴定和分离可能在 ESCC 的癌症管理方案的改进中发挥重要作用。此外，CSC 可能被用作 ESCC 化放疗反应的预测工具。不同的方法，如体外功能测定、使用各种细胞内和细胞表面标志物的细胞分选以及异种移植技术，经常用于不同癌症，包括 ESCC 中 CSCs 的鉴定和分离。然而，这些方法都不能单独保证 CSC 群体的完全分离。因此，采用多种方法结合用于可靠地检测和分离 CSCs。在此，我们描述了通过 Hoechst 33342 染色后细胞分选，然后进行体外功能测定和体内小鼠异种移植技术，从 ESCC 细胞中鉴定和分离 CSCs 的方法。
