Stimulation of endogenous kinin receptors promotes growth of androgen-independent prostate cancer PC3 cells via activation of the mitogenic extracellular-signal-regulated kinase (ERK) pathway. In the present study, we show that kinin-mediated mitogenic signalling and prostate-cell growth involves two subtypes of bradykinin (BK) receptors, B1R and B2R. Specific stimulation of either B1R or B2R by their respective agonists des-Arg9-BK and Lys-BK promoted ERK activation and cell growth, whereas selective blockade with specific antagonists des-Arg9-[Leu8]BK and Hoe 140 respectively obliterated this effect, indicating the presence of both receptor subtypes. However, blockade of B1R also inhibited B2R-mediated ERK activation and cell growth, and, similarly, antagonism of B2R inhibited the B1R-mediated response. Furthermore, both B1R and B2R agonists promoted internalization of B1R, whereas both receptor antagonists blocked this effect. The B1R ligands des-Arg9-BK and des-Arg9-[Leu8]BK had no effect on the binding of BK to B2R, as demonstrated by radioligand competitive binding studies. However, blockade of either B1R or B2R impaired the ability of the reciprocal receptor to produce inositol phosphates, suggesting that the interaction between B1R and B2R is proximal to activation of phospholipase C. These results provide evidence for the existence of B1R–B2R complexes in prostate cancer PC3 cells and demonstrate that antagonism of one receptor interferes with the signalling ability of the other, possibly at the level of receptor–Gαq protein coupling. Selective inhibition of B1R, which is up-regulated in injured and cancerous tissue, may be beneficial for the treatment of advanced prostate cancer.
Abbreviations used: AT1R, angiotensin II type 1 receptor; BK, bradykinin; B1R, BK receptor 1; B2R, BK receptor 2; EGF, epidermal growth factor; ERK, extracellular-signal-regulated kinase; GPCR, G-protein-coupled receptor; MAPK, mitogen-activated protein kinase.
Present address: Wallenberg Neuroscience Center, Institute of Physiological Sciences, SE-22184 Lund, Sweden.