基于流式细胞术的方案，使用 Wnt/β-catenin 信号通路报告基因评估结直肠肿瘤干细胞的细胞毒性和克隆形成能力。

FACS-based protocol to assess cytotoxicity and clonogenic potential of colorectal cancer stem cells using a Wnt/β-catenin signaling pathway reporter.

机构信息

Department of Surgical Oncological and Stomatological Sciences, University of Palermo, Palermo 90127, Italy.

Laboratory for Experimental Oncology and Radiobiology, Center for Experimental and Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, Amsterdam 1105 AZ, the Netherlands.

出版信息

STAR Protoc. 2021 Oct 19;2(4):100880. doi: 10.1016/j.xpro.2021.100880. eCollection 2021 Dec 17.

DOI:10.1016/j.xpro.2021.100880

PMID:34712995

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8529548/

Abstract

Cancer stem cells (CSCs) play a key role in tumor initiation and progression. A real-time tool to evaluate the activation of CSC-specific signaling pathways is crucial for the study of this cancer cell subset. Here, we present a protocol to monitor, , the activation of Wnt/β-catenin signaling pathway, which is considered a functional biomarker for colorectal CSCs (CR-CSCs). This flow-cytometry-based protocol allows it to isolate CR-CSCs and to evaluate their cytotoxicity upon anti-tumor treatments. For complete details on the use and execution of this protocol, please refer to Di Franco et al. (2021).

摘要

癌症干细胞 (CSCs) 在肿瘤的发生和发展中起着关键作用。一种实时评估 CSC 特异性信号通路激活的工具对于研究这种癌细胞亚群至关重要。在这里，我们提出了一种监测结直肠 CSCs（CR-CSCs）中 Wnt/β-连环蛋白信号通路激活的方案。该方案基于流式细胞术，可用于分离 CR-CSCs，并评估它们在抗肿瘤治疗后的细胞毒性。有关该方案使用和执行的完整详细信息，请参阅 Di Franco 等人（2021 年）。