Free calcium and calmodulin levels in acinar carcinoma and normal acinar cells of rat pancreas

Abstract

Exposure of acinar carcinoma cells and normal acinar cells of rat pancreas to the muscarinic agonist drug carbamylcholine stimulated 45Ca2+ outflux from 45Ca2+-labeled cells. More rapid outflux of 45Ca2+ was detected for carcinoma cells following muscarinic stimulation than for normal cells. Direct fluorometric measurement of cytosolic Ca2+ under basal (unstimulated) conditions in quin 2-loaded cells revealed significantly lower concentration of free Ca2+ in carcinoma cells (approximately 180 nM) than in normal cells (approximately 200 nM). Stimulation with 1 mM carbamylcholine increased the cytosolic Ca2+ concentration in carcinoma and normal cells to approximately 1900 nM, after which carcinoma cells removed cytosolic Ca2+ at a faster rate to a post-stimulation plateau concentration of approximately 140 nM, in comparison to normal cells which obtained a post-stimulation plateau concentration of approximately 300 nM. Essentially identical differences between carcinoma and normal cells were detected upon stimulation with the peptidergic agonist cholecystokinin octapeptide. Finally, carcinoma cells demonstrated approximately 3 times greater calmodulin concentration than normal acinar cells. Also, the calmodulin antagonist drug W7 (N-6-(aminohexyl)-5-chloro-1-naphthalene sulfonamide) inhibited the carbamylcholine-induced release of intracellular Ca2+ in acinar carcinoma cells. These results indicate that neoplastic pancreatic acinar cells have retained mechanisms of muscarinic- and peptidergic-stimulated intracellular Ca2+ release, and implicate calmodulin as a regulatory factor in secretagogue activation of intracellular Ca2+ release. We propose that the more rapid decline of intracellular Ca2+ concentration following muscarinic or peptidergic stimulation and the increased intracellular calmodulin concentration indicate calmodulin-mediated down-regulation of free cytosolic Ca2+ in acinar carcinoma cells to levels lower than those of normal cells.