HGF-Induced DU145 Cell Scatter Assay

Abstract

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.