The effects of bombesin on polyphosphoinositide and calcium metabolism in Swiss 3T3 cells

Abstract

Bombesin, a peptide mitogen for a variety of cell types, acts as a typical Ca2+-mobilizing hormone in Swiss 3T3 fibroblasts. At its mitogenic concentrations (1-25 nM), bombesin stimulates polyphosphoinositide turnover, i.e. breakdown of phosphatidylinositol 4,5-bisphosphate and a concomitant increase in inositol phosphates in a time- and dose-dependent manner. In particular, bombesin induces an initial transient increase in inositol 1,4,5-trisphosphate concentration, followed by an increase in the concentration of inositol 1,3,4-trisphosphate. Also, within 30 s of bombesin addition, the mass of 1,2-diacylglycerol nearly doubles and remains at this level for up to 60 min. Intracellular [Ca2+] measurements with a photoprotein, aequorin, demonstrate that bombesin stimulates a transient rise in cytosolic free Ca2+ concentration. A mobilization of Ca2+ from an intracellular pool is observed as a dose-dependent, transient increase in 45Ca2+ efflux from prelabeled cells, both in the presence and absence of extracellular Ca2+. Bombesin also induces a sustained increase in Ca2+ influx rate and stimulates 3-O-methyl-D-glucose transport across the plasma membrane. These composite results indicate that the mitogenic effect of bombesin is mediated through an activation of the Ca2+ messenger system.