Epidermal growth factor increases phosphoinositide turnover and intracellular free calcium in an immortalized human myometrial cell line independent of the arachidonic acid metabolic pathway

OBJECTIVES: The objectives of this study were to determine whether epidermal growth factor increases intracellular calcium and phosphoinositide turnover in human myometrial cells by a tyrosine kinase-mediated mechanism, to evaluate an obligatory role for arachidonic acid metabolites in these actions, and to compare the actions of epidermal growth factor and oxytocin. STUDY DESIGN: Intracellular calcium and phosphoinositide turnover were measured in a myometrial cell line after stimulation with epidermal growth factor (0.1 to 100 nmol/L) or oxytocin (20 nmol/L). The effects of nifedipine, thapsigargin, genestein and tyrphostin, the guanosine triphosphate binding protein antagonist GPA-7, indomethacin, and nordihydroguaiaretic acid were determined. Data were analyzed by analysis of variance and Duncan's multiple-range test. RESULTS: Epidermal growth factor stimulated phosphoinositide turnover and increased intracellular calcium in a dose-dependent manner (median effective concentration 2.6 nmol/L). In contrast to oxytocin, the effects of epidermal growth factor were inhibited by tyrosine kinase inhibitors but not by GPA-7. Indomethacin and nordihydroguaiaretic acid did not inhibit the epidermal growth factor-stimulated increase in intracellular calcium. CONCLUSIONS: The acute epidermal growth factor-stimulated increase in intracellular calcium in this myometrial cell line is primarily derived from release of calcium from intracellular stores, and it involves the activation of a tyrosine kinase, presumably the epidermal growth factor receptor. Arachidonic acid metabolites are not obligatory intermediates. Oxytocin increases phosphoinositide turnover and intracellular calcium by a distinctly different pathway.