Human submandibular gland (HSG) cell line as a model for studying salivary gland Ca2+signalling mechanisms

Abstract

The human submandibular gland cell line (HSG) has been used as a model for studying the molecular mechanisms of salivary cells. The aim of this study was to investigate some aspects of salivary Ca2+ signalling. We focused on the presence and function of specific molecular markers of salivary cells to see whether this cell line retained normal salivary characteristics, despite the neoplastic changes. We detected the M3 acetylcholine receptor and intracellular salivary amylase mRNA with RT-PCR. Carbachol treatment caused a rapid, transient elevation of [Ca2+]i, showing that the cholinergic receptors are functional in HSG cells. Protein kinase C activation by phorbol-esther PMA, prior to carbachol treatment, inhibited the normal Ca2+ signalling pathway in HSG cells. Using selective antagonists, we also identified the dominant muscarinic receptor subtype M3 on HSG cells. We also observed that functional extracellular purinergic receptors were present on HSG cells and coupled to intracellular Ca2+ signalling. Our results suggested that the coupling mechanisms of these receptors remained relatively intact despite the neoplastic transformation. This enables us to use this cell line to model the role of muscarinic and purinergic control of salivary gland function, cell proliferation and differentiation.