We employed RNA interference to determine the role of tissue transglutaminase II (TGase II) in motility of cancer cells. Down-regulation of TGase II by small interfering RNA against TGase II impaired adhesion and motility of HeLa cells by decreasing phosphorylation of the protein kinase, Akt, and reactive oxygen species (ROS). Over-expression of TGase II showed opposite effects. These results suggest potential utility of TGase II for development of therapeutic anti cancer vaccine.