Fram Sally T, Wells Claire M, Jones Gareth E
Randall Division of Cell and Molecular Biophysics, King’s College London, London, UK.
Methods Mol Biol. 2011;769:31-40. doi: 10.1007/978-1-61779-207-6_3.
Epithelial mesenchymal transition (EMT) is a multi-stage process whereby epithelial cells lose their cell:cell adhesions and acquire the capacity to migrate independently. It is a process that is important in normal development and is thought to be adopted by some invasive cancer cells. EMT requires modifications in cell shape and substratum adhesions and these events are dependent on the reorganisation of the actin cytoskeleton. Hepatocyte growth factor (HGF) is a mitogenic growth factor that is well known to induce such a conversion, termed "cell scattering", in Madin Darby canine kidney (MDCK) cells. Recently, we have developed an alternative model of cell scattering using the human prostate cancer cell line, DU145. Like MDCK cells, DU145 cells normally grow as tight colonies with firm cell:cell junctions, but they can be induced to 'scatter' upon HGF stimulation. Here, we describe the optimised protocol for conducting and analysing an HGF-induced DU145 scatter assay. This model is particularly useful for monitoring changes in actin cytoskeletal organisation and dynamics, cell:cell adhesions, and cell migration in human cells that respond to HGF stimulation.
上皮-间质转化（EMT）是一个多阶段过程，在此过程中上皮细胞失去细胞间黏附，并获得独立迁移的能力。这一过程在正常发育中很重要，并且被认为某些侵袭性癌细胞也会经历。EMT需要细胞形状和基质黏附的改变，而这些事件依赖于肌动蛋白细胞骨架的重组。肝细胞生长因子（HGF）是一种促有丝分裂生长因子，众所周知它能在Madin Darby犬肾（MDCK）细胞中诱导这种转变，即所谓的“细胞散射”。最近，我们利用人前列腺癌细胞系DU145开发了一种细胞散射的替代模型。与MDCK细胞一样，DU145细胞通常紧密聚集生长，细胞间连接牢固，但在HGF刺激下它们可被诱导“散射”。在此，我们描述了进行和分析HGF诱导的DU145散射实验的优化方案。该模型对于监测响应HGF刺激的人细胞中肌动蛋白细胞骨架组织和动力学、细胞间黏附以及细胞迁移的变化特别有用。
