Histone deacetylase 6 (HDAC6) promotes cell motility and contributes to the metastasis of cancers. The purpose of this study was to investigate the role of HDAC6 in human bladder cancer for the first time. The results showed that HDAC6 promotes the migration and invasion of bladder cancer cells by targeting the cytoskeletal protein cortactin. Furthermore, a colony formation assay as well as in vitro migration and invasion assays demonstrated that this migration and invasion was suppressed by the HDAC6-specific inhibitor tubacin. In addition, cortactin is the substrate of SIRT2, which also belongs to the family of histone deacetylases. We demonstrated that by using SIRT2-specific siRNA combined with tubacin treatment, the cell migratory and invasive abilities were dramatically suppressed. Taken together, we conclude that HDAC6 and SIRT2 work synergistically to promote cell migration and invasion in bladder cancer, and the HDAC6-specific inhibitor tubacin may be regarded as a novel therapeutic agent for bladder cancer.