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Growth factor regulation of the amylase promoter in a differentiating salivary acinar cell line
Author(s) -
Zheng Changyu,
Hoffman Matthew P.,
McMillan Teresa,
Kleinman Hynda K.,
O'Connell Brian C.
Publication year - 1998
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/(sici)1097-4652(199812)177:4<628::aid-jcp13>3.0.co;2-l
Subject(s) - matrigel , acinar cell , ductal cells , epidermal growth factor , biology , growth factor , cell culture , microbiology and biotechnology , hepatocyte growth factor , luciferase , chemistry , transfection , endocrinology , pancreas , biochemistry , genetics , receptor
Salivary glands contain two major epithelial cell types: acinar cells which produce the primary salivary secretion, including amylase, and ductal cells which reabsorb electrolytes but also secrete kallikrein. Here we investigated salivary acinar cell differentiation in vitro using the activity of the salivary amylase and tissue kallikrein promoters as markers of acinar cell and ductal cell differentiation, respectively. Each of the promoter sequences was cloned into a replication‐deficient adenoviral vector containing the luciferase reporter gene. Previous studies showed that a human submandibular gland cell line (HSG) differentiated into acinar cells when cultured on a reconstituted basement membrane matrix (Matrigel). The luciferase activity of the amylase promoter vector (AdAMY‐luc) was low in HSG cells cultured on plastic, where they grow as an epithelial monolayer. The promoter activity increased approximately tenfold when HSG cells were cultured on Matrigel and developed an acinar phenotype. Under the same conditions, the luciferase activity of the kallikrein promoter (AdKALL‐luc) was not induced. Because HSG cells demonstrate acinar cell morphology, but not amylase gene expression, when cultured on laminin‐1, certain soluble components of Matrigel were tested for their ability to induce the amylase promoter during in vitro differentiation of acinar cells. We find that epidermal growth factor (EGF) and transforming growth factor‐alpha (TGF‐α), which are present in the basement membrane, and hepatocyte growth factor (HGF) increase activity of the amylase promoter. Other basement membrane‐derived growth factors such as TGF‐β, basic fibroblast growth factor (bFGF), and platelet‐derived growth factor (PGDF), as well as tumor necrosis factor (TNF‐α), keratinocyte growth factor (KGH), nerve growth factor (NGF) and interferon gamma (IFN‐γ) were inactive. This system will be further exploited to study the mechanisms by which extracellular matrix molecules and growth factors regulate salivary acinar cell differentiation. J Cell Physiol 177:628–635, 1998. Published 1998 Wiley‐Liss, Inc.