Department of Metabolism, Development and Reproduction, Institute of Reproductive and Developmental Biology (IRDB), Imperial College London, Du Cane Road, W12 0HS, UK.
Department of Biological and Medical Sciences, Faculty of Health and Life Sciences, Oxford Brookes University, Gipsy Lane, Headington, Oxford, OX3 0BP, UK.
Acta Histochem. 2020 Dec;122(8):151654. doi: 10.1016/j.acthis.2020.151654. Epub 2020 Nov 3.
While metastasis - the spread of cancer from the primary location to distant sites in the body - remains the principle cause of cancer death, it is incompletely understood. It is a complex process, requiring the metastatically successful cancer cell to negotiate a formidable series of interconnected steps, which are described in this paper. For each step, we review the range of in vitro assays that may be used to study them. We also provide a range of detailed, step-by-step protocols that can be undertaken in most modestly-equipped laboratories, including methods for converting qualitative observations into quantitative data for analysis. Assays include: (1) a gelatin degradation assay to study the ability of endothelial cells to degrade extracellular matrix during tumour angiogenesis; (2) the morphological characterisation of cells undergoing epithelial-mesenchymal transition (EMT) as they acquire motility; (3) a 'scratch' or 'wound-healing' assay to study cancer cell migration; (4) a transwell assay to study cancer cell invasion through extracellular matrix; and (5) a static adhesion assay to examine cancer cell interactions with, and adhesion to, endothelial monolayers. This toolkit of protocols will enable researchers who are interested in metastasis to begin to focus on defined aspects of the process. It is only by further understanding this complex, fascinating and clinically relevant series of events that we may ultimately devise ways of better treating, or even preventing, cancer metastasis. The assays may also be of more broad interest to researchers interested in studying aspects of cellular behaviour in relation to other developmental and disease processes.
虽然转移——癌症从原发部位扩散到身体远处的部位——仍然是癌症死亡的主要原因，但它并没有被完全理解。这是一个复杂的过程，需要转移性成功的癌细胞通过一系列艰巨的相互关联的步骤，本文描述了这些步骤。对于每一步，我们回顾了可能用于研究这些步骤的一系列体外检测方法。我们还提供了一系列详细的、逐步的方案，这些方案可以在大多数设备不太齐全的实验室中进行，包括将定性观察转化为用于分析的定量数据的方法。检测方法包括：(1)明胶降解检测，用于研究肿瘤血管生成过程中内皮细胞降解细胞外基质的能力；(2)对经历上皮-间充质转化(EMT)的细胞进行形态学特征分析，因为它们获得了运动性；(3)“划痕”或“伤口愈合”检测，用于研究癌细胞迁移；(4)Transwell 检测，用于研究癌细胞穿过细胞外基质的侵袭；(5)静态粘附检测，用于检查癌细胞与内皮单层的相互作用和粘附。这个方案工具包将使有兴趣研究转移的研究人员能够开始专注于该过程的特定方面。只有通过进一步了解这一系列复杂、有趣且与临床相关的事件，我们才有可能最终设计出更好地治疗甚至预防癌症转移的方法。这些检测方法可能也会引起对研究与其他发育和疾病过程相关的细胞行为方面感兴趣的研究人员的广泛关注。
