Three-Dimensional Quantification of Spheroid Degradation-Dependent Invasion and Invadopodia Formation.
Three-Dimensional Quantification of Spheroid Degradation-Dependent Invasion and Invadopodia Formation.
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DOI：
10.1186/s12575-018-0085-6
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发表时间：
2018
影响因子：
6.4
通讯作者：
Magalhaes M
中科院分区：
文献类型：
--
作者：
Goertzen C;Eymael D;Magalhaes M
侵足是一种富含肌动蛋白的蛋白水解结构，使癌细胞能够侵入周围组织。几种体外侵袭试验已经在文献中使用，从定向定量分析到复杂的三维（3D）分析。这些检测的主要局限性之一是在模拟肿瘤微环境的三维（3D）环境中缺乏可量化的降解依赖性入侵。在这篇文章中，我们描述了一种基于目前可用的肿瘤球体模型的新的侵袭和降解实验，它允许肿瘤的长期高分辨率成像，精确的量化，以及基质降解和多通道免疫细胞化学的可视化。通过将降解标记物（DQ-Green BSA）结合到基底膜基质中，我们证明了在3D中量化癌细胞诱导的基质降解的能力。此外，我们描述了一种技术，以产生肿瘤球体的组织学切片，允许在三维肿瘤球体中检测侵足形成。这项新技术为在体外研究癌症提供了明显的优势，并将有助于解决有关癌细胞入侵动力学的关键问题。
Invadopodia are actin-rich, proteolytic structures that enable cancer cell to invade into the surrounding tissues. Several in vitro invasion assays have been used in the literature ranging from directional quantitative assays to complex three-dimensional (3D) analyses. One of the main limitations of these assays is the lack of quantifiable degradation-dependent invasion in a three-dimensional (3D) environment that mimics the tumor microenvironment. In this article, we describe a new invasion and degradation assay based on the currently available tumor spheroid model that allows long-term high-resolution imaging of the tumor, precise quantification, and visualization of matrix degradation and multichannel immunocytochemistry. By incorporating a degradation marker (DQ-Green BSA) into a basement-membrane matrix, we demonstrate the ability to quantitate cancer cell-induced matrix degradation in 3D. Also, we describe a technique to generate histological sections of the tumor spheroid allowing the detection of invadopodia formation in the 3D tumor spheroid. This new technique provides a clear advantage for studying cancer in vitro and will help address critical questions regarding the dynamics of cancer cell invasion.
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影响因子：
4.6
作者：
Zanoni M;Piccinini F;Arienti C;Zamagni A;Santi S;Polico R;Bevilacqua A;Tesei A
通讯作者：
Tesei A
DOI：
10.1083/jcb.201103045
发表时间：
2011-11-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
Magalhaes MA;Larson DR;Mader CC;Bravo-Cordero JJ;Gil-Henn H;Oser M;Chen X;Koleske AJ;Condeelis J
通讯作者：
Condeelis J
影响因子：
4.8
作者：
Debnath, J;Muthuswamy, SK;Brugge, JS
通讯作者：
Brugge, JS
DOI：
10.3791/52686
发表时间：
2015-05-01
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Vinci M;Box C;Eccles SA
通讯作者：
Eccles SA
影响因子：
4.8
作者：
Goertzen, Cameron G.;Dragan, Magdalena;Bhattacharya, Moshmi
通讯作者：
Bhattacharya, Moshmi