Analysis of Histone Deacetylase-Dependent Effects on Cell Migration Using the Stripe Assay.
Analysis of Histone Deacetylase-Dependent Effects on Cell Migration Using the Stripe Assay.
复制标题
使用条纹测定分析组蛋白脱乙酰酶依赖性对细胞迁移的影响
DOI：
10.1007/978-1-4939-6527-4_5
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发表时间：
2017
影响因子：
--
通讯作者：
Thanos S
中科院分区：
文献类型：
--
作者：
Mertsch S;Thanos S
对于正常的胚胎发育/形态发生，细胞迁移和归巢是精心策划的重要事件，需要特定的细胞机制。在诸如癌症的疾病中，失调的细胞迁移是一个主要问题。因此，许多努力正在进行中，以了解肿瘤细胞迁移的分子机制，并产生更有效的肿瘤治疗。细胞迁移测定是最常用的功能测定之一。伤口愈合试验或Boyden室试验是这些试验的变体。几乎所有这些都是二维测定，并且细胞一次只能在一个基底上迁移。这与细胞同时面对不同表面并与不同细胞类型相互作用的体内情况相反。为了接近这种体内情况，我们使用了Bonhoeffer及其同事设计的条纹试验的修改版本来检查轴突引导的机制。该试验的设计允许细胞在同时提供的两种不同底物之间做出决定。利用交替的神经元基板迁移分析，我们可以部分模拟复杂的脑肿瘤细胞在体内的情况。在这里，我们描述了详细的协议进行修改版本的条纹试验，以观察底物依赖性的迁移效应在体外，分析的影响Rho依赖性激酶（ROCKS），组蛋白脱乙酰酶（HDAC）和其他分子对胶质瘤细胞。
For normal embryonic development/morphogenesis, cell migration and homing are well-orchestrated and important events requiring specific cellular mechanisms. In diseases such as cancer deregulated cell migration represents a major problem. Therefore, numerous efforts are under way to understand the molecular mechanisms of tumor cell migration and to generate more efficient tumor therapies. Cell migration assays are one of the most commonly used functional assays. The wound-healing assay or the Boyden chamber assay are variations of these assays. Nearly all of them are two-dimensional assays and the cells can only migrate on one substrate at a time. This is in contrast to the in vivo situation where the cells are faced simultaneously with different surfaces and interact with different cell types. To approach this in vivo situation we used a modified version of the stripe assay designed by Bonhoeffer and colleagues to examine mechanisms of axonal guidance. The design of this assay allows cells to decide between two different substrates offered at the same time. Utilizing alternating neuronal substrates for migration analyses we can partially mimic the complex in vivo situation for brain tumor cells. Here we describe the detailed protocol to perform a modified version of the stripe assay in order to observe substrate-dependent migration effects in vitro, to analyze the effect of Rho-dependent kinases (ROCKS), of histone deacetylases (HDACs) and of other molecules on glioma cells.
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影响因子：
6.1
作者：
Wang, C. Joanne;Li, Xiong;Levchenko, Andre
通讯作者：
Levchenko, Andre
影响因子：
3.6
作者：
MAXWELL, GD
通讯作者：
MAXWELL, GD
影响因子：
3.9
作者：
Wang, Chenlu;Chowdhury, Sagar;Losert, Wolfgang
通讯作者：
Losert, Wolfgang
影响因子：
12.7
作者：
Louis DN;Ohgaki H;Wiestler OD;Cavenee WK;Burger PC;Jouvet A;Scheithauer BW;Kleihues P
通讯作者：
Kleihues P