Department of Cell Biology, Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Sci Signal. 2012 Jul 3;5(231):rs5. doi: 10.1126/scisignal.2002677.
The scattering of cultured epithelial cells in response to hepatocyte growth factor (HGF) is a model system that recapitulates key features of metastatic cell behavior in vitro, including disruption of cell-cell adhesions and induction of cell migration. We have developed image analysis tools that do not require fluorescence tagging and that automatically track and characterize three aspects of scattering in live cells: increase in cell motility, loss of cell-cell adhesion, and spatial dispersion of cells (the redistribution of cells during scattering). We used these tools to screen a library of drugs, and we identified several efficient inhibitors of scattering, which we classified as selective inhibitors of either motility or loss of cell-cell adhesion, or as nonselective inhibitors. We validated the inhibitors and putative targets from this screen in two unrelated model cell lines. Using pharmacological treatments and RNA interference (RNAi), we found that nonsteroidal anti-inflammatory drugs inhibited cell-cell dissociation, that indirubins inhibited cell motility, and that cyclin-dependent kinase 1 and ribosomal S6 kinase were signaling intermediates in HGF-induced cell scattering. This assay is suitable for larger-scale screenings of chemical compounds or RNAi libraries.
细胞培养上皮细胞对肝细胞生长因子（HGF）的散射是一个模型系统，该系统体外再现了转移性细胞行为的关键特征，包括破坏细胞-细胞黏附以及诱导细胞迁移。我们开发了不需要荧光标记的图像分析工具，并能够自动跟踪和描述活细胞散射的三个方面：细胞迁移能力增加、细胞-细胞黏附丧失以及细胞空间分散（细胞在散射过程中的重新分布）。我们使用这些工具筛选了一个药物文库，发现了几种有效的散射抑制剂，我们将其分类为运动或细胞-细胞黏附丧失的选择性抑制剂，或非选择性抑制剂。我们在两种不相关的模型细胞系中验证了该筛选的抑制剂和潜在靶点。通过药理学处理和 RNA 干扰（RNAi），我们发现非甾体抗炎药抑制细胞分离，吲哚布芬抑制细胞运动，细胞周期蛋白依赖性激酶 1 和核糖体 S6 激酶是 HGF 诱导的细胞散射的信号中间物。该测定适用于化学化合物或 RNAi 文库的大规模筛选。
