Several physiological parameters were examined for inducing acinar cell proliferation and corresponding increased expression of β1-4 galactosyltransferase. In this study, dietary changes causing acinar cell proliferation included the following: the introduction of animals to a liquid diet (causing gland atrophy) followed by re-introduction of solid chow, gustatory stimulation provided by the introduction of 0.5% citric acid to animal drinking water, and removal of the submandibular gland with subsequent reliance on the parotid gland for saliva protein and fluid. Alterations in growth factor levels were produced by injecting animals with a chronic (three-day) regimen of either nerve growth factor (NGF) or epidermal growth factor (EGF). In all cases of acinar cell proliferation in vivo, generated by the above treatments, cell-surface galactosyltransferase was detected along with the unique expression of a 4.5-kb proliferation-associated mRNA. Parotid gland proliferation could be blocked in all cases by the injection of the galactosyltransferase specific modifier protein, a-lactalbumin. Propranolol, a β-adrenergic receptor antagonist, blocked proliferation in all cases except EGF treatment. EGFinduced proliferation could, however, be prevented if the animals were treated with monoclonal antibody to EGF receptor or with the galactosyltransferase modifier a-lactalbumin.
As a comparison, human parotid tissue samples obtained from neoplastic pleomorphic adenomas, mucoepidermoid carcinoma, adenoid cystic carcinoma, and a bulimia patient were analyzed for galactosyltransferase expression by Northern blot of mRNA and plasma membrane isolation. Elevated levels of galactosyltransferase were found in all neoplastic tissue preparations as well as in the bulimia sample. Amylase synthesis was reduced in samples compared with surrounding normal tissue from the same patient. In vitro cell culturing of pleomorphic adenoma cells in the presence of galactosyltransferase modifier a-lactalbumin and substrate UDP-galactose inhibited proliferation in a dose-dependent fashion. Southern blot analysis of DNA from neoplastic parotid cells showed an alteration in chromosomal gene structure for the galactosyltransferase activator cDNA from the adenoid cystic carcinoma.
These results for induced acinar cell proliferation as well as human neoplastic pathologies suggest a direct role for cell surface β1-4 galactosyltransferase in signaling growth. Furthermore, the proliferation-associated activity of galactosyltransferase suggests that it may be considered as a new type of cell growth regulator.
Get full access to this article
View all access options for this article.
References
1.
Arvan, P. and Castle, J.D. (1982): Plasma Membrane of the Rat Parotid Gland: Preparation and Partial Characterization of a Fraction Containing the Secretory Surface , J Cell Biol95: 8-19.
2.
Banya, E.N.; Runyan, R.B.; Scully, N.F.; Reichner, J.; Lopez, L.C.; and Shur, B.D. (1986): Cell Surface Galactosyltransferase as a Recognition Molecule During Development, Molec Cell Biochem72: 141-151.
3.
Barka, T. (1965): Induced Cell Proliferation: the Effect of Isoproterenol , Exp Cell Res37: 662-679.
4.
Bernacki, R.J. and Kim, U. (1977): Concomitant Elevations in Serum Sialyltransferase Activity and Sialic Acid Content in Rats with Metastasizing Mammary Tumors , Science195: 577-580.
5.
Bernfeld, P. (1955): Rapid Assay of a-Amylase Enzyme Activity, Meth Enzymol1 : 149.
6.
Bosmann, H.B. and Hall, T.C. (1974): Enzyme Activity in Invasive Tumors of Human Breast and Colon, Proc Natl Acad Sci USA71: 1833-1837.
7.
Brown-Grant, K. (1961): Enlargement of Salivary Glands in Mice Treated with Isopropylnoradrenaline, Nature191: 1076-1078.
8.
Carlin, C.R.; Tollefson, A.E.; Brady, H.A.; Hoffman, B.L.; and Wold, W.S.M. (1989): Epidermal Growth Factor Receptor is Down-regulated by a 10,400 MW Protein Encoded by the E3 Region of Adenovirus, Cell 57: 135-144.
9.
Chang, A.; Todd, R.; Chow, P.; McBride, J.; Chiang, T.; Matossian, K.; Gallagher, G.; Donoff, B.; Shklar, G.; and Wong, D. (1989): TGF-Alpha and EGF receptor mRNAs in Human Oral Cancer, J Dent Res68: 292, Abst. No. 883.
10.
Chen, W.S.; Lazar, C.S.; Poene, M.; Tsien, R.Y.; Gill, G.N.; and Rosenfeld, M.G. (1987): Requirement for Intrinsic Protein Tyrosine Kinase in the Immediate and Late Actions of EGF Receptor, Nature328: 820-823.
11.
Cleveland, D.W.; Lopata, M.A.; MacDonald, R.J.; Cowan, N.J.; Rutter, W.J.; and Kirschner, M.W. (1980): Number and Evolutionary Conservation of a- and β-Tubulin and Cytoplasmic β- and γ-Actin Genes Using Specific Cloned cDNA Probes , Cell20: 95-105.
12.
Cowan, N.J.; Wilde, C.D.; Chow, L.T.; and Wefald, F.C. (1981): Structural Variation Among Human β-Tubulin Genes, Proc Natl Acad Sci USA78: 4877-4881.
13.
Fairbanks, G.; Steck, T.L.; and Wallach, D.E. (1971): Electrophoretic Analysis of Major Polypeptides of the Erythrocyte Membrane, Biochemistry10: 2606-2617.
14.
Fujii, M.; Shalloway, D.; and Verma, 1. ( 1989): Gene Regulation by Tyrosine Kinases: src Protein Activates Various Promotors, Including c-fos, Molec Cell Biol9: 2493-2499.
15.
Greene, L.A. and Shooter, E.M. (1980): The Nerve Growth Factor: Biochemistry, Synthesis and Mechanism of Action, Ann Rev Neuroscience3: 353-402.
16.
Hall, H.D. and Schneyer, C.A. (1973): Role of Autonomic Pathways in Disuse Atrophy of Rat Parotid, Proc Soc Exp Biol Med143: 19-22.
17.
Hall, H.D. and Schneyer, C.A. (1977): Functional Mediation of Compensatory Enlargement of Parotid, Cell Tissue Res184: 249-254.
18.
Hall, H.D. and Schneyer, C.A. (1978): Neural Regulation of Compensatory Enlargement of the Parotid Gland of the Rat, Cell Tissue Res187: 147-151.
19.
Humphreys-Beher, M.G. (1989): Restoration of a-Lactalbumin-inhibited Rat Parotid Gland Hypertrophy and Hyperplasia by Agents Specific for Membrane Glycoprotein N-Acetylglucosamine, Arch Oral Biol34: 811-819.
20.
Humphreys-Beher, M.G.; Bunnell, B.; Ledbetter, D.H.; VAN Tuinen, P.; and Kidd, V.J. (1986): Molecular Cloning and Chromosomal Localization of Human 4β-Galactosyltransferase, Proc Natl Acad Sci USA83: 8918-8923. Correction in PNAS 86: 8747 (1989).
21.
Humphreys-Beher, M.G.; Imell, M.; Jentoft, N.; Gleason, M.; and Carlson, D.M. (1984): Isolation and Characterization of UDP-galactose: N-acetylglucosamine 4β-Galactosyltransferase Activity Induced in Rat Parotid Glands Treated with Isoproterenol, J Biol Chem259: 5799-5802.
22.
Humphreys-Beher, M.G. and Schneyer, C.A. (1987): Cell Surface Expression of 4β-Galactosyltransferase Accompanies Rat Parotid Gland Hypertrophy Induced by Changes in Diet, Biochem J246: 387-391.
23.
Humphreys-Beher, M.G.; Schneyer, C.A.; Kidd, V.J.; and Marchase, R.B. (1987): Isoproterenol-mediated Parotid Gland Hypertrophy is Inhibited by Effectors of 4β-Galactosyltransferase, J Biol Chem262: 11706-11713.
24.
Humphreys-Beher, M.G. and Wells, D.J. (1984): Metachromatic Staining Patterns of Basic Proline-rich Proteins from Human and Rat Saliva in Sodium Dodecyl-Sulfate-Polyacrylamide Gels, J Appl Biochem6: 353-360.
25.
Inoue, H.; Kikuchi, K.; and Nishino, M. (1986): Effects of Epidermal Growth Factor on the Synthesis of DNA and Polyamine in Isoproterenol-stimulated Murine Parotid Gland, J Biochem100: 605-613.
26.
Ip, C. and Dao, T.L. (1977): Increase in Serum and Tissue Glycosyltransferases and Glycosidases in Tumor-bearing Rats, Cancer Res37: 3442-3447.
27.
Johnson, D.A. (1983): Differences in Basic Proline-rich Proteins in Rat Saliva Following Chronic Isoproterenol Treatment or Maintenance on a Liquid Diet, Arch Oral Biol28: 549-554.
28.
Kidd, V.J.; Wallace, R.B.; Itakura, K.; and Woo, S.L.C. (1983): α1-Antitrypsin Deficiency Detection by Direct Analysis of the Mutation in the Gene, Nature304: 230-234.
29.
Klohs, W.D.; Willson, J.R.; and Weiser, M.M. (1982): UDP-galactose Inhibition of BALB/3 T12 Cell Growth , Exp Cell Res141: 365-374.
30.
Kornfeld, R. and Kornfeld, S. (1985): Assembly of Asparagine-linked Oligosaccharides , Ann Rev Biochem54: 631-664.
31.
Krstulovic, A.M. (1982): Investigations of Catecholamine Metabolism Using High Performance Liquid Chromatography, J Chromatog229: 1-34.
32.
Lamont, J.F.; Gammon, M.T.; and Isselbacher, K.J. (1977): Cell-surface Glycosyltransferases in Cultured Fibroblasts. Increased Activity and Release During Serum Stimulation of Growth, Proc Natl Acad Sci USA74: 1086-1090.
33.
Levin, P.A.; Falko, J.M.; Dixon, K.; Gallup, E.M.; and Saunders, W. (1980): Benign Parotid Enlargement in Bulimia, Ann Int Med93: 827-829.
34.
Loken, M.R. and Stall, A.M. (1982): Flow Cytometry as an Analytical and Preparative Tool in Immunology, J Immunol Meth50: R85-R112.
35.
Lopez, L.C. and Shur, B.D. (1988): Comparison of Two Independent cDNA Clones Reported to Encode β1,4 Galactosyltransferase, Biochem Biophys Res Commun156: 1223-1229.
36.
Luo, W.; Kidd, V.J.; Humphreys-Beher, M.G.; Slavkin, H.C.; and Snead, M.L. (1987): Expression of 4β-Galactosyltransferase mRNA in the Ectoderm-derived Cell Transition to Growth, J Cell Biol105: 49a, Abst. 265.
37.
Marchase, R.B.; Kidd, V.J.; Rivera, A.A.; and Humphreys-Beher, M.G. (1988): Cell-surface Expression of 4β-Galactosyltransferase Accompanies Rat Parotid Acinar Cell Transition to Growth, J Cell Biochem36: 453-465.
38.
Morley, D.J. and Hodes, M.E. (1988): Amylase Expression in Human Parotid Neoplasms: Evidence by in situ Hybridization for a Lack of Transcription of the Amylase Gene, J Histochem Cytochem36: 487-491.
39.
Muenzer, J.; Bildstein, C.; Gleason, M.; and Carlson, D.M. (1979): Purification of Proline-rich Proteins from Parotid Glands of Isoproterenol-treated Rats, J Biol Chem254: 5623-5628.
Oliver, C.; Waters, J.F.; Tolbert, C.L.; and Kleinman, H.K. (1987): Growth of Exocrine Acinar Cells on a Reconstituted Basement Membrane Gel, In Vitro Cell Dev Biol23: 465-473.
42.
Patt, L.M.; Van Nest, G.A.; and Grimes, W.J. (1975): A Comparison of Glycosyltransferase Activities and Malignant Properties in Normal and Transformed Cells Derived from BALB/c Mice, Cancer Res35: 438-441.
43.
Pierce, M.; Turley, E.A.; and Roth, S. (1980): Cell Surface Galactosyltransferase Activities. In: International Review of Cytology, G.H. Bourne, J.F. Danielli, and K.W. Jean, Eds., New York: Academic Press , pp. 2-44.
44.
Podolsky, D.K. and Isselbacher, K.J. (1984): Characterization of Monoclonal Antibodies to Serum Galactosyltransferase, Proc Natl Acad Sci USA81: 2529-2533.
45.
Podolsky, D.K. and Weiser, M.M. (1975): Galactosyltransferase Activities in Human Sera: Detection of a Cancer-associated Isoenzyme, Biochem Biophys Res Commun65: 545-551.
46.
Podolsky, D.K. and Weiser, M.M. (1979): Detection, Purification and Characterization of a Human Cancer-associated Galactosyltransferase Acceptor, Biochem J178: 279-287.
47.
Podolsky, D.K.; Weiser, M.M.; and Isselbacher, K.J. (1978): Inhibition of Growth of Transformed Cells and Tumors by an Endogenous Acceptor of Galactosyltransferase , Proc Natl Acad Sci USA75: 4426-4430.
48.
Pugsley, A.P. and Schnaitman, C.A. (1979): Factors Affecting the Electrophoretic Mobility of the Major Outer Membrane Proteins of Escherichia coli in Polyacrylamide Gels, Biochim Biophys Acta581: 163-178.
49.
Rauch, S.D. and Herzog, D.B. (1987): Parotidectomy for Bulimia : A Dissenting View , Am J Otolaryngol8: 376-380.
50.
Roseman, S. (1970): The Synthesis of Complex Carbohydrates by Multiglycosyltransferase Systems and Their Potential Function in Intercellular Adhesion, Chem Phys Lipids5: 270-274.
51.
Roth, J.; Lentze, M.J.; and Berger, E.G. (1985): Immunocytochemical Demonstration of Ecto-Galactosyltransferase in Absorptive Intestinal Cells, J Cell Biol100 : 118-125.
52.
Roth, S.; McGuire, E.J.; and Roseman, S. (1971): Evidence for Cell Surface Glycosyltransferase , J Cell Biol51: 536-547.
53.
Roth, S.; Roelke, M.; and Dorsey, J. (1977): Cell Surface Glycosyltransferase on Malignant and Nonmalignant Cells. In: Growth, Kinetics and Biochemical Regulation of Normal and Malignant Cells, B. Drewinko and R.M. Humphrey, Eds., Baltimore : Williams and Wilkins, pp. 245-259.
54.
Roth, S. and White, D. (1972): Intercellular Contact and Cell-surface Galactosyltransferase Activity, Proc Natl Acad Sci USA69: 485-489.
55.
Schaterle, G.E. and Pollach, R.L. (1973): A Simplified Method for the Quantified Assay of Small Amounts of Protein in Biologic Material, Anal Biochem51: 654-656.
56.
Schneyer, C.A. (1962): Salivary Gland Changes After Isoproterenol-induced Enlargement, Am J Physiol203: 232-246.
57.
Schneyer, C.A. (1972): Regulation of Salivary Gland Size. In: Regulation of Organ and Tissue Growth, R.J. Goss, Ed., New York: Academic Press, pp. 211-232.
58.
Schneyer, C.A. and Humphreys-Beher, M.G. (1988): Effects of Epidermal Growth Factor and Nerve Growth Factor on Isoproterenol-induced DNA Synthesis in Rat Parotid and Pancreas Following Removal of Submandibular-Sublingual Glands, J Oral Pathol17: 250-256.
59.
Schneyer, C.A. and Humphreys-Beher, M.G. (1989): Inhibition of Isoproterenol-induced DNA and RNA Synthesis in Rat Parotid and Pancreas Following Removal of the Submandibular-Sublingual Glands, Cell Tissue Res256: 361-363.
60.
Selye, H.; Cantin, M.; and Veilleux, R. (1961): Abnormal Growth and Sclerosis of the Salivary Glands Induced by Chronic Treatment with Isoproterenol, Growth25: 243-248.
61.
Shur, B.D. (1984): The Receptor Function of Galactosyltransferase During Cellular Interactions, Molec Cell Biochem61: 143-156.
62.
Thomas, P.S. (1980): Hybridization of Denatured RNA and Small DNA Fragments Transferred to Nitrocellulose, Proc Natl Acad Sci USA77: 5201-5205.
63.
Towbin, H.; Staehelin, T.; and Gordon, J. (1979): Electrophoretic Transfer of Proteins from Polyacrylamide Gels to Nitrocellulose Sheets: Procedure and Some Applications, Proc Natl Acad Sci USA76: 4350-4354.
64.
Zelles, T.; Blazsek, J.; Ko'Bor, A.; and Gelencser, F. (1984): A Glandula Parotis Gusztatorikus Ingerrel Letrehozott Megnagyobbada sa, Fogovosoi Szemle77: 315-318 (in Hungarian).
