Major role for active extension in the formation of processes by ras-transformed fibroblasts

Expression of constitutively active Ras protein in fibroblasts results in enhanced cell motility, invasion competence and morphological changes including the formation of elongate cellular processes. These processes have been shown to resemble retraction tails formed passively behind nontransformed cells by movement relative to sites of cell-substrate attachment. However, analysis presented here reveals that active extension mechanisms also play a role in the formation of these processes. Extension of distal process ends occurs at 0.42+/-0.44 microm/min in ras-transformed fibroblasts and accounts for 63.6+/-27.5% of observed process lengths. Active process extension by ras-transformed fibroblasts also persists in the presence of cell-cell contacts. Studies conducted using actin or microtubule antagonists, and correlation of process behavior followed by fixation and immunostaining reveal that process extension requires intact actin and microtubule networks. Other analyses reveal that active extension plays a significantly smaller role in the formation of processes by non-transformed control fibroblasts. These observations demonstrate that constitutively active Ras enhances process extension in fibroblasts and is a causal factor in process extension by fibroblasts in the presence of cell-cell contacts. Moreover, these studies demonstrate that process extension by ras-transformed fibroblasts is accomplished through mechanisms similar to those thought to drive active extension of processes by other cell types including neurons. These findings suggest that extension of cellular processes could play an important role in the metastatic behavior of ras-transformed fibroblasts as well as the response of untransformed fibroblasts to receptor mediated signal transduction events.