通过有丝分裂摇脱和显微镜检查对关卡适应进行实验测定

Experimental Determination of Checkpoint Adaptation by Mitotic Shake-Off and Microscopy.

作者信息

Swift Lucy H, Golsteyn Roy M

机构信息

POETIC Laboratory for Preclinical and Drug Discovery Studies, Departments of Pediatrics and Oncology, Alberta Children's Hospital Research Institute, Arnie Charbonneau Cancer Institute, University of Calgary, Calgary, AB, Canada.

Cancer Cell Laboratory, Department of Biological Sciences, University of Lethbridge, Lethbridge, AB, Canada.

出版信息

Methods Mol Biol. 2018;1769:159-168. doi: 10.1007/978-1-4939-7780-2_10.

DOI:10.1007/978-1-4939-7780-2_10

PMID:29564823

Abstract

Cells that undergo checkpoint adaptation arrest at and then abrogate the G2/M cell cycle checkpoint to enter mitosis with damaged DNA. Cells surviving this process frequently contain micronuclei, which can lead to genomic change and chromothripsis. In this chapter we describe how to induce checkpoint adaptation and detect it by time-lapse video and immunofluorescence microscopy and how to isolate cells undergoing checkpoint adaptation from a total cell population.

摘要

经历检查点适应的细胞会在G2/M细胞周期检查点处停滞，然后解除该检查点，从而带着受损的DNA进入有丝分裂。在此过程中存活下来的细胞通常含有微核，这可能导致基因组改变和染色体碎裂。在本章中，我们将描述如何诱导检查点适应，以及如何通过延时视频和免疫荧光显微镜检测检查点适应，还将介绍如何从整个细胞群体中分离出正在经历检查点适应的细胞。