Oral squamous cell carcinoma (OSCC) is the most malignant lesion occurring in the head and neck. The Rho-kinases (ROCKs), effectors of Rho proteins, are involved in actin cytoskeletal organization, cell migration, and maintenance cortex. The HA-1077 inhibits the ROCKs. This study aimed to evaluate the effect of treatment with HA-1077 on cell motility in SCC-4 cells, a cell line originating from human OSCC. F-actin of SCC-4 cells treated or not with HA-1077 (1, 50 and 100mol/l), and also HA-1077 50mol/l and/or inhibitors Y-27632 30mol/l was stained with rhodamine-conjugated phalloidin and analyzed by confocal microscopy. Approximately 110cells/well, control and treated with HA-1077 (25, 50, and 100mol/l) were added to the migration plate assay. In addition, 110cells/well, control and treated with HA-1077 50mol/l, were tested by invasion assays (plate coated with Matrigel). The inhibition of ROCKs with HA-1077 and/or Y-27632 leads to morphological changes, affecting the organization of the actin. The inhibitory effect of HA-1077 (P<0.0001) was dose dependent as the number of cells migrated at 100mol/l was statistically different: 25mol/l (P<0.0001) and 50mol/l (P<0.01). The number of cells treated with HA-1077 50mol/l decreased compared with control cells that invaded through Matrigel (P<0.0001). This study shows an inhibitory effect of HA-1077 on cell migration and invasion, suggesting that the use of HA-1077 can be a potential therapy for OSCC.