October 1995
Volume 36, Issue 11
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Articles  |   October 1995
Thrombin increases expression of urokinase receptor by activation of the thrombin receptor.
Author Affiliations
  • Z Yang
    Cancer Research Institute, University of California at San Francisco 94143-0128, USA.
  • R L Cohen
    Cancer Research Institute, University of California at San Francisco 94143-0128, USA.
  • G M Lui
    Cancer Research Institute, University of California at San Francisco 94143-0128, USA.
  • D A Lawrence
    Cancer Research Institute, University of California at San Francisco 94143-0128, USA.
  • M A Shuman
    Cancer Research Institute, University of California at San Francisco 94143-0128, USA.
Investigative Ophthalmology & Visual Science October 1995, Vol.36, 2254-2261. doi:
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    • Get Citation

      Z Yang, R L Cohen, G M Lui, D A Lawrence, M A Shuman; Thrombin increases expression of urokinase receptor by activation of the thrombin receptor.. Invest. Ophthalmol. Vis. Sci. 1995;36(11):2254-2261.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: To investigate the effect of thrombin on the urokinase plasminogen activator receptor (u-PAR) in retinal pigment epithelial (RPE) cells. METHODS: The authors analyzed u-PAR mRNA by Northern blot hybridization. Retinal pigment epithelial cell surface u-PAR was assayed by measuring the amount of functional urokinase plasminogen activator (u-PA) bound to cells at saturation. Retinal pigment epithelial cells were derived from fetal retinal tissue and established in primary cell culture. RESULTS: Thrombin increased u-PAR mRNA 4-fold in RPE cells examined by Northern blot hybridization, whereas the amount of thrombin receptor mRNA was unchanged. Thrombin stimulated u-PA binding to RPE cells 2.5- to 5-fold in a time- and dose-dependent manner. Hirudin, a thrombin antagonist, completely blocked the effects of thrombin on u-PAR expression in RPE cells. Phosphatidylinositol phospholipase C treatment of RPE cells resulted in the abolition of thrombin-induced u-PA binding. Recombinant soluble u-PAR competitively inhibited two-chain u-PA binding to the surface of thrombin-treated RPE cells. A thrombin receptor agonist peptide (SFLLRNPNDKYEPF) also induced a 2.5-fold increase in binding of u-PA to the surface of RPE cells. CONCLUSION: Thrombin increases u-PAR expression by RPE cells by a mechanism involving activation of the seven transmembrane thrombin receptor.

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