Sui Xin, Geng Jian-Hao, Yu Hui-Ming, Li Yong-Heng, Wang Wei-Hu
Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Radiation Oncology, Peking University Cancer Hospital and Institute.
Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Radiation Oncology, Peking University Cancer Hospital and Institute;
J Vis Exp. 2019 Aug 21(150). doi: 10.3791/60046.
The presence of cancer stem cells (CSCs) has been associated with relapse or poor outcomes after radiotherapy. Studying radioresistant CSCs may provide clues to overcoming radioresistance. Voltage-gated calcium channel α2δ1 subunit isoform 5 has been reported as a marker for radioresistant CSCs in non-small cell lung cancer (NSCLC) cell lines. Using calcium channel α2δ1 subunit as an example of a CSC marker, methods to study the radiosensitivity of CSCs in NSCLC cell lines are presented. CSCs are sorted with putative markers by flow cytometry, and the self-renewal capacity of sorted cells is evaluated by sphere formation assay. Colony formation assay, which determines how many cells lose the ability to generate descendants forming the colony after a certain dose of radiation, is then performed to assess the radiosensitivity of sorted cells. This manuscript provides initial steps for studying the radiosensitivity of CSCs, which establishes the basis for further understanding of the underlying mechanisms.
癌症干细胞（CSCs）的存在与放疗后的复发或不良预后相关。研究放射抗性CSCs可能为克服放射抗性提供线索。电压门控钙通道α2δ1亚基异构体5已被报道为非小细胞肺癌（NSCLC）细胞系中放射抗性CSCs的标志物。以钙通道α2δ1亚基作为CSC标志物的示例，介绍了研究NSCLC细胞系中CSCs放射敏感性的方法。通过流式细胞术用假定的标志物对CSCs进行分选，并通过成球试验评估分选细胞的自我更新能力。然后进行集落形成试验，该试验确定在一定剂量的辐射后有多少细胞失去产生形成集落的后代的能力，以评估分选细胞的放射敏感性。本手稿提供了研究CSCs放射敏感性的初步步骤，为进一步了解其潜在机制奠定了基础。
