Translationally Controlled Tumor-associated Protein (TCTP) plays a role in a plethora of normal and cancer cell functions including cell cycle progression, cell growth and metastasis. Our previous studies showed that TCTP interacts with cellular cytoskeleton and is localized, in cell-type specific manner, on actin filaments in various types of ovarian cancer cells. Here we used small interfering RNA (siRNA) for silencing TCTP expression in human ovarian surface epithelial noncancerous cell line HIO180, ovarian carcinoma cell lines SKOV3 and OVCAR3 and analyzed effect of TCTP silencing on actin cytoskeleton and cell motility. We show that a down regulation of TCTP caused dramatic restructuring and redistribution of actin filaments in HIO180, SKOV3 and OVCAR3 cells and resulted in cell motility increase. This previously unidentified dependence of actin cytoskeleton remodeling and cell motility on TCTP level might be responsible for high metastatic potential and aggressiveness of ovarian cancer cells and will help to pinpoint novel targets for anticancer therapies..