Spoerri Loredana, Beaumont Kimberley A, Anfosso Andrea, Haass Nikolas K
The University of Queensland Diamantina Institute, Translational Research Institute, The University of Queensland, 37 Kent St, Woolloongabba, Brisbane, QLD, 4102, Australia.
The Centenary Institute, Newtown, NSW, Australia.
Methods Mol Biol. 2017;1612:401-416. doi: 10.1007/978-1-4939-7021-6_29.
Aberrant cell cycle progression is a hallmark of solid tumors; therefore, cell cycle analysis is an invaluable technique to study cancer cell biology. However, cell cycle progression has been most commonly assessed by methods that are limited to temporal snapshots or that lack spatial information. Here, we describe a technique that allows spatiotemporal real-time tracking of cell cycle progression of individual cells in a multicellular context. The power of this system lies in the use of 3D melanoma spheroids generated from melanoma cells engineered with the fluorescent ubiquitination-based cell cycle indicator (FUCCI). This technique allows us to gain further and more detailed insight into several relevant aspects of solid cancer cell biology, such as tumor growth, proliferation, invasion, and drug sensitivity.
异常的细胞周期进程是实体瘤的一个标志；因此，细胞周期分析是研究癌细胞生物学的一项非常有价值的技术。然而，细胞周期进程最常用的评估方法仅限于时间快照或缺乏空间信息。在这里，我们描述了一种技术，该技术能够在多细胞环境中对单个细胞的细胞周期进程进行时空实时追踪。该系统的强大之处在于使用了由基于荧光泛素化的细胞周期指示剂（FUCCI）工程改造的黑色素瘤细胞生成的三维黑色素瘤球体。这项技术使我们能够更深入、更详细地了解实体癌细胞生物学的几个相关方面，如肿瘤生长、增殖、侵袭和药物敏感性。
