title: Regulation of scu-PA secretion and u-PA receptor expression in osteoblast-like cells.
abstract: The production of proteolytic enzymes by osteoblasts is considered important for initiating osteoclastic bone resorption. Using the established cell line NY as an example of osteoblast-like cells, the effect of intracellular cyclic AMP (cAMP) and protein kinase C (PKC) on plasminogen activator secretion and its specific binding to the cells were investigated. HT-1080 cells were used as the control. NY cells predominantly secrete single-chain urokinase-type plasminogen activator (scu-PA) and some two-chain u-PA. Both scu-PA and u-PA were present in the cell surface and cell lysate of NY cells, and their distribution in HT-1080 cells was quite similar to that of NY cells. Exposing cells to phorbol myristate acetate (PMA) or dibutyryl cyclic AMP (db cAMP) enhanced the secretion of scu-PA and two-chain u-PA, whereas 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H7) decreased scu-PA secretion, indicating that it is enhanced by protein kinase C (PKC) as well as by cAMP in NY cells. On the other hand, in HT-1080 cells, PMA decreased the level of two-chain u-PA secretion into the conditioned medium. The binding assay of 125I-DFP-u-PA to NY cells revealed the presence of a single class of binding sites with a Kd of 2.23 nM and Bmax of 0.82 x 10(6) binding sites/cell. PMA however, altered neither the Kd nor the Bmax. Dibutyryl cAMP increased the Bmax 1.9 fold. Thus, NY cells secrete u-PA and express specific binding sites on the cell surface, which are modulated by cAMP and PKC.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Physiology, Kinki University of Medicine, Osakasayama, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nonaka
authors.forename: K
authors.initials: K
authors.lastname: Okada
authors.forename: H
authors.initials: H
authors.lastname: Fukao
authors.forename: S
authors.initials: S
authors.lastname: Ueshima
authors.forename: H
authors.initials: H
authors.lastname: Kikuchi
authors.forename: S
authors.initials: S
authors.lastname: Tanaka
authors.forename: O
authors.initials: O
authors.lastname: Matsuo
mesh_terms: D000242:Cyclic AMP / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D010006:Osteoblasts / Q000378:metabolism*; D011493:Protein Kinase C / Q000502:physiology; D011956:Receptors, Cell Surface / Q000096:biosynthesis*; D055293:Receptors, Urokinase Plasminogen Activator; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
chemical_list: C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D000242:Cyclic AMP; D011493:Protein Kinase C; D014568:Urokinase-Type Plasminogen Activator