用于趋化因子受体拮抗剂的琼脂糖斑点趋化性测定法。
An agarose spot chemotaxis assay for chemokine receptor antagonists.
作者信息
Vinader Victoria, Al-Saraireh Yousef, Wiggins Helen L, Rappoport Joshua Z, Shnyder Steve D, Patterson Laurence H, Afarinkia Kamyar
机构信息
The Institute of Cancer Therapeutics, University of Bradford, West Yorkshire BD7 1DP, UK.
出版信息
J Pharmacol Toxicol Methods. 2011 Nov-Dec;64(3):213-6. doi: 10.1016/j.vascn.2011.01.004. Epub 2011 Feb 1.
INTRODUCTION
Chemokines are important players in directing the migration of cancer cells as part of the metastatic process. The aim of this study is to develop an easy-to-perform, reliable, and inexpensive assay for rapid analysis of anti-chemotactic activity of chemokine antagonists under a number of experimental conditions.
METHODS
An agarose spot containing the chemokine chemoattractant is applied to a glass petri dish. Live cells in a media, both with and without a chemokine antagonist, are added to the dish and, following cell adhesion, the migration under the agarose spot is observed and analysed by microscopy.
RESULTS
In the absence of CXCL12 in the agarose, no migration under the agarose spot is detected. In the presence of CXCL12, significant migration under the agarose spot is observed which can be retarded if a neutralising monoclonal antibody or a small molecule antagonist is added to the media.
DISCUSSION
This experimental configuration is a reliable, inexpensive and easy-to-perform chemotaxis assay, which enables assessment of the activity of CXCR4 antagonists.
引言
趋化因子在引导癌细胞迁移作为转移过程的一部分中起着重要作用。本研究的目的是开发一种易于操作、可靠且廉价的检测方法，用于在多种实验条件下快速分析趋化因子拮抗剂的抗趋化活性。
方法
将含有趋化因子化学引诱剂的琼脂糖斑点应用于玻璃培养皿。将含有或不含有趋化因子拮抗剂的培养基中的活细胞添加到培养皿中，细胞黏附后，通过显微镜观察并分析琼脂糖斑点下的细胞迁移情况。
结果
在琼脂糖中不存在CXCL12的情况下，未检测到琼脂糖斑点下的细胞迁移。在存在CXCL12的情况下，观察到琼脂糖斑点下有明显的细胞迁移，如果向培养基中添加中和性单克隆抗体或小分子拮抗剂，这种迁移会受到抑制。
讨论
这种实验配置是一种可靠、廉价且易于操作的趋化性检测方法，能够评估CXCR4拮抗剂的活性。
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