title: beta-Adrenergic-responsive activation of extracellular signal-regulated protein kinases in salivary cells: role of epidermal growth factor receptor and cAMP.
abstract: The beta-adrenergic receptor agonist isoproterenol exerts growth-promoting effects on salivary glands. In this study, activation of ERKs, members of the mitogen-activated protein kinase family, by isoproterenol was examined in a human salivary gland cell line (HSY). Immunoblot analysis indicated that isoproterenol (10(-5) M) induced transient activation of ERK1/2 (4.4-fold relative to basal at 10 min) similar to that caused by EGF (6.7 fold). Isoproterenol, like EGF, also induced phosphorylation of the EGF receptor. However, inhibition of EGF receptor phosphorylation by the tyrphostin AG-1478 only partially attenuated isoproterenol-induced ERK phosphorylation, whereas EGF-responsive ERK activation was completely blocked. The G(i) inhibitor pertussis toxin also caused partial inhibition of isoproterenol-stimulated ERK activation. The cAMP analog 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (CPT-cAMP) and the cAMP-elevating agents IBMX and cholera toxin produced transient ERK1/2 activation, similar to the effect of isoproterenol, in HSY cells. The stimulatory effects of isoproterenol and cAMP on ERK phosphorylation were not reduced by the PKA inhibitor H-89, whereas the Src family inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidase (PP2) and transfection of a dominant-negative Src construct diminished isoproterenol-induced ERK activation. Isoproterenol induced marked overexpression of the cell growth-related adhesion molecule CD44, and this effect of isoproterenol was abolished by the ERK pathway inhibitor PD-98059. In summary, we show a dual mechanism of isoproterenol-induced ERK phosphorylation in HSY cells-one pathway mediated by EGF receptor transactivation and the other by an EGF receptor-independent pathway possibly mediated by cAMP. Our results also suggest that isoproterenol-induced growth of salivary tissue may involve ERK-mediated CD44 expression.
authors.affiliation: Geriatric Research, Education and Clinical Center, South Texas Veterans Health Care System, 7400 Merton Minter Blvd., San Antonio, TX 78229-4404, USA.
authors.forename: Chih-Ko
authors.initials: CK
authors.lastname: Yeh
authors.forename: Paramita M
authors.initials: PM
authors.lastname: Ghosh
authors.forename: Howard
authors.initials: H
authors.lastname: Dang
authors.forename: Qun
authors.initials: Q
authors.lastname: Liu
authors.forename: Alan L
authors.initials: AL
authors.lastname: Lin
authors.forename: Bin-Xian
authors.initials: BX
authors.lastname: Zhang
authors.forename: Michael S
authors.initials: MS
authors.lastname: Katz
mesh_terms: D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D002460:Cell Line; D000242:Cyclic AMP / Q000502:physiology*; D004789:Enzyme Activation; D066246:ErbB Receptors / Q000502:physiology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006801:Humans; D018960:Hyaluronan Receptors / Q000502:physiology; D007545:Isoproterenol / Q000494:pharmacology*; D012469:Salivary Glands / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*
chemical_list: D000318:Adrenergic beta-Agonists; D018960:Hyaluronan Receptors; D000242:Cyclic AMP; D066246:ErbB Receptors; D048049:Extracellular Signal-Regulated MAP Kinases; D007545:Isoproterenol