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The Ras-homology (Rho) GTPases were initially linked to regulation of actin cytoskeletal architecture, maintenance of cell shape and polarity, and to cell migration. Subsequent studies have shown that Rho GTPases are involved in highly diverse cellular and biological functions, many of which contribute to tumorigenesis. Here, we first present an overview of the regulation and more ‘traditional’ functions of Rho GTPases. We then discuss the consequences of deregulated Rho GTPase signaling during cancer initiation and development, and some recent developments connecting Rho GTPases to metabolic reprogramming and to the generation of cancer cell microvesicles.