Department of Pharmacology, University of California, San Diego, La Jolla, CA, USA.
Moores Cancer Center, Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA.
Methods Mol Biol. 2021;2294:151-163. doi: 10.1007/978-1-0716-1350-4_11.
During the metastatic process, carcinoma cells form invadopodia, F-actin enriched protrusive structures, to degrade the extracellular matrix (ECM) in order to invade the surrounding stroma and intravasate into the circulatory system. In this chapter, we describe the 2D-fluorescent matrix degradation assay, a highly sensitive and reproducible in vitro method used to measure invadopodia-mediated ECM degradation. We provide a detailed protocol on how to prepare the glass coverslips with fluorescent gelatin matrix and a standardized method to quantify gelatin degradation and invadopodia formation in order to evaluate cell invasion.
在转移过程中，癌细胞形成侵袭伪足，富含 F-肌动蛋白的伸出结构，降解细胞外基质（ECM），以便侵袭周围基质并浸润循环系统。在本章中，我们描述了二维荧光基质降解测定法，这是一种高度敏感且可重复的体外方法，用于测量侵袭伪足介导的 ECM 降解。我们提供了一个详细的方案，说明如何制备带有荧光明胶基质的玻璃盖玻片，以及一种标准化的方法来量化明胶降解和侵袭伪足形成，以评估细胞侵袭。
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