The polymerization of microfilaments and their subsequent rearrangements under the control of actin-myosin interactions are two major processes that underlie the morphogenetic reactions of cells. We studied their role in the spreading of normal and transformed REF52tetRas fibroblasts with adjustable ras-oncogene expression. Treatment with inhibitors of cell contractility (Y27632 or blebbistatin) led to the disappearance of actin bundles and focal adhesions; however, pseudopodial activity in both normal and transformed cells remained high. Under these conditions, spreading was more accelerated in normal cells then in ras-transformed cells. In normal cells treated with low concentrations of latrunculin A actin polymerization was suppressed, stress fibers and focal adhesions were preserved, but lamellipodial activity was lost and spreading was dramatically inhibited. In transformed fibroblasts treated with low doses of latrunculin, actin bundles and focal adhesions almost disappeared, but pseudopodial activity was apparent and spreading was less suppressed. Therefore, the most significant process in the regulation of cell spreading and polarization is the microfilament polymerization at the leading edge. ras-Transformed cells are less sensitive to inhibitors that affecting the cytoskeletal structure than nontransformed cells. Possible mechanisms that underlie the difference are discussed.