Urokinase (uPA) was originally identified in human urine for its ability to catalyse the transformation of plasminogen into its active form, plasmin which degrades fibrin and extracellular matrix components. Two major, functionally independent regions have been identified in the uPA molecule: a non-catalytic N-terminal region (residues 1-135) and a large catalytic region (residues 159-411) spaced by the "connecting peptide" (residues 136-158). Binding of uPA to its specific surface receptor (uPAR) amplifies cell surface plasminogen activation, thus enhancing pericellular proteolysis. The uPAR, linked to the lipid bilayer via a glycosylphosphatidylinositol anchor, mediates signaling through the assembly of a multiprotein complex with transmembrane receptors, like the integrins, EGFR, GPCRs. Receptor engagement with uPA results in a variety of cell responses, including increased proliferation, survival, migration and invasion. These responses may be enhanced by the concomitant binding of the uPA "connecting peptide" region to αvβ5 integrin, thus favoring uPAR-integrin association. Receptors engaged with uPA exhibit a high affinity for vitronectin, stimulating cell adhesion. The uPA/uPAR system is regarded as one of the key systems driving tumour invasion and metastases. Different strategies to prevent the activity of the protease, as well as the interactions of uPAR with integrins and GPCRs have been designed. Many preclinical studies are ongoing and, at least, two uPA-related compounds have reached Phase II clinical trials. The aim of this review is to provide a comprehensive picture of the functionally relevant interactions, together with a description of the promising compounds and strategies to control uPA activity and signaling in human pathologies.