Abstract
Tissue non-specific alkaline phosphatase is a membrane-bound glycoprotein enzyme which is characterized by its phosphohydrolytic, protein phosphatase, and phosphotransferase activities. This enzyme is distributed virtually in all mammalian tissues, particularly during embryonic development. Its expression is stagespecific and can be demonstrated in the developing embryo as early as the 2-cell stage. It has been suggested that tissue non-specific alkaline phosphatase might play a role in tissue formation. In the study reported here, a genetransfer approach was employed to investigate possible roles for this enzyme by inserting the cDNA for rat tissue non-specific alkaline phosphatase into CHO and LLC-PK1 cells. Permanently transfected cell-lines expressing varying levels of alkaline phosphatase were estblished. The data showed that functional enzyme was expressed in the transfected cells. Cell spreading and attachment were enhanced in transfected CHO cells expressing high levels of tissue non-specific alkaline phosphatase but not in the LLC-PK1 cells. Further, in CHO cells, proliferation was shown to be inversely proportional to the level of the tissue non-specific alkaline phosphatase expression. Homotypic cell association was demonstrated in both alkaline phosphatase-positive and alkaline phosphatase-negative cells in both CHO and LLC-PK1 celllines. Taken together, these findings suggest that in addition to a role in mineralization of bone, tissue nonspecific alkaline phosphatase might also play a role in other cell activities, including those related to differentiation, such as cell-cell or cell-substrate interaction and proliferation.
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References
Andracchi S, Korte GF (1991) Expression of plasma membrane alkaline phosphatase in normal and regenerating choriocapillaris in the rabbit. Acta Anat (Basel) 141:289–293
Beliveau R, Fortier G, Vachon V (1988) Effect of magnesium on ATP labelling by kidney brush border membrane. Int J Biochem 20:375–380
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein binding. Anal Biochem 72:248–254
Carpenter G (1981) Vanadate, epidermal growth factor, and the stimulation of DNA synthesis. Biochem Biophys Res Commun 102:1115–1121
Farley JR, Puzas JE, Baylink DJ (1982) Effect of skeletal alkaline phosphatase inhibitors on bone cell proliferation in vitro. Miner Electrolyte Metab 7:316–323
Fedarko NS, Bianco P, Vetter U, Robey PG (1990) Human bone cell enzyme expression and cellular heterogeneity correlation of alkaline phosphatase enzyme activity with cell cycle. J Cell Physiol 144:115–121
Feldbush TL, Lafrenz D (1991) Alkaline phosphatase on activated B cells characterization of the expression of alkaline phosphatase on activated B cells. J Immunol 147:3690–3695
Giancotti FG, Ruoslahti E (1990) Elevated levels of the alpha-5 beta-1 fibronectin receptor suppressed the transformed phenotype of Chinese hamster ovary cells. Cell 60:849–859
Hahnel AC, Schultz GA (1989) Cloning and characterization of a cDNA encoding alkaline phosphatase in mouse embryonal carcinoma cells. Clin Chim Acta 186:174–174
Habnel AC, Rappolee A, Millan JL, Manes T, Ziomek CA, Theodosiou NG, Werb Z, Pedersen RA, Schultz GA (1990) Two alkaline phosphatase genes are expressed during early development in the mouse embryo. Development 110:555–564
Hanrylak K, Kihn L, Rutkowski D, Stinson RA (1989) Purified tetrameric alkaline phosphatase: the effect of treatments with phosphatidylinositol phospholipase C and sodium sulfate. Clin Chim Acta 186:197–202
Harris H (1989) The human alkaline phosphatase: what we know and what we don't know. Clin Chim Acta 186:133–150
Hillman JR, Seliger WG, Epling GP (1975) Histochemistry and ultrastructure of adrenal cortical development in the golden hamster. Gen Comp Endocrinol 25:14–16
Hirst R, Horwitz A, Buck C, Rohrschneider L (1986) Phosphorylation of the fibronectin receptor complex in cells transformed by oncogenes that encode tyrosine kinase. Proc Natl Acad Sci USA 83:6470–6474
Karczmar AG, Berg GG (1951) Alkaline phosphatase during limb development and regeneration of Amblystoma opacum and Amblystoma punctatum. J Exp Zool 117:139–144
Majeska RG, Nair BC, Rodan GA (1985) Glucocorticoid regulation of alkaline phosphatase in the osteoblastic osteosarcoma cell line ROS 17/2.8. Endocrinology 116:170–178
McComb RB, Bowers GN, Posen S (1979) Alkaline phosphatase. Plenum Press, New York
McCulloch CAG, Fair CA, Tenenbaum HC, Limeback H, Homareau R (1990) Clonal distribution of osteoprogenitor cells in cultured chick periostea: functional relationship to bone formation. Dev Biol 140:352–361
McWhinnie DJ, Saunders JW (1966) Developmental patterns and specificities of alkaline phosphatase in the embryonic chick limb. Dev Biol 14:169–173
Muller K, Schellenberger V, Borneleit P, Treide A (1991) The alkaline phosphatase from bone: transphosphorylating activity and kinetic mechanism. Biochim Biophys Acta 1076:308–313
Nakamura T, Nakamura K, Stinson RA (1988) Release of alkaline phosphatase from human osteosarcoma cells by phosphatidylinositol phospholipase C: effect of tunicamycin. Arch Biochem Biophys 265:190–196
Owen TA, Aronow M, Shalhoub V, Barone LM, Wilming L, Tassinari MS, Kennedy MB, Pockwnse S, Lan JB, Stein GS (1990) Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. J Cell Physiol 143:420–430
Pizauro JM, Ciancaglini P, Leone FA (1992) Phosphotransferase activity associated with rat osseous plate alkaline phosphatase: a possible role in biomineralization. Int J Biochem 24:1391–1396
Puzas JE, Brand JS (1982) Complex and unexpected effects of vanadate ion on bone cell proliferation. Calcif Tissue Int 34:S13
Reeves ME (1992) A metastatic tumor cell line has greatly reduced levels of a specific homotypic cell adhesion molecule activity. Cancer Res 52:1546–1552
Sarrouilhe D, Lalegerie P, Baudry M (1992) Endogenous phosphorylation and dephosphorylation of rat liver plasma membrane proteins, suggesting a 18 kDa phosphoprotein as a potential substrate for alkaline phosphatase. Biochim Biophys Acta 1118:116–122
Stinson RA, McPhee JL, Colier HB (1987) Phosphotransferase activity of human alkaline phosphatase and role of enzymeZn2+. Biochim Biophys Acta 913:272–278
Thiede MA, Yoon K, Golub EE, Noda M, Rodan GA (1988) Structure and expression of rat osteosarcoma (Ros 17/2.8) alkaline phosphatase: product of a single copy gene. Proc Natl Acad Sci USA 85:319–323
Weiss MJ, Henthorn PS, Lafferty MA, Slaughtie C, Raducha M, Harris H (1986) Isolation and characterization of a cDNA encoding a human liver/bone/kidney-type alkaline phosphatase. Proc Natl Acad Sci USA 83:7182–7186
Whitt MA, Buonocore L, Rose JK (1991) TransfectACE-TM reagent: Transient transfection frequencies>90%. Focus 13:8–12
Yamaguchi Y, Ruoslahti E (1988) Expression of human proteoglycan in Chinese hamster ovary cells inhibits cell proliferation. Nature 336:244–246
Yoon K, Golub E, Rodan GA (1989) Alkaline phosphatase cDNA transfected cells promote calcium and phosphate deposition. Connect Tissue Res 22:17–25
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Hui, M., Hu, M. & Tenenbaum, H.C. Changes in cell adhesion and cell proliferation are associated with expression of tissue non-specific alkaline phosphatase. Cell Tissue Res 274, 429–437 (1993). https://doi.org/10.1007/BF00314539
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DOI: https://doi.org/10.1007/BF00314539