Invasion and metastasis models for studying RhoGDI2 in bladder cancer

Invasion and metastasis are the critical steps in cancer progression that lead to death from this disease. Intense investigation into the underlying mechanisms of metastasis has revealed a complex set of signaling pathways that regulate the process. Since the mid-1980s, it has been demonstrated that the Rho family of proteins plays a major role in these pathways. Proteins that regulate Rho, including guanine nucleotide exchange factors, GTPase-activating proteins, and Rho GDP dissociation inhibitors (RhoGDIs), have also been shown to contribute to cancer progression. Among this group of Rho-regulating proteins is RhoGDI2 (RhoGDIbeta/LyGDI/GDID4/RabGDIbeta). Our laboratory initially identified RhoGDI2 as a metastasis suppressor due to its differential expression between metastatically capable and poorly metastatic bladder cancer cell lines. Over the subsequent years, in vivo and in vitro systems have been used to model steps in the metastatic cascade and to test how the expression of RhoGDI2 affected those processes. This chapter describes several of the more significant methods used to investigate the role of RhoGDI2 in bladder cancer invasion and metastasis. These methods include an in vitro assay for invasion using bladder organ cultures, lung metastasis assays in immunocompromised murine hosts, polymerase chain reaction-based quantification of metastatic burden, and derivation of increasingly metastatic cell lines.