Urokinase Activity in Corneal Fibroblasts may be Modulated by DNA Damage and Secreted Proteins(para)
Proteases like urokinase-type plasminogen activator (uPA) play an important role in tumor invasion. Cells derived from ultraviolet radiation (UVR)-induced corneal sarcomas of Monodelphis domestica produce relatively high levels of uPA compared to the untransformed keratocytes suggesting a mechanism for their invasiveness. Because UVR is known to stimulate uPA production in many cell types, UVR exposure may further increase uPA expression in corneal tumor cells, thus enhancing their ability to infiltrate. We investigated control of basal uPA levels and the induction of uPA by UVR in transformed and untransformed corneal keratocytes from Monodelphis. These studies took advantage of the fact that Monodelphis possesses an active photolyase that can be stimulated to remove UVR-induced pyrimidine dimers by exposure to longwavelength visible photoreactivating light (PRL). Our studies showed that significant induction of uPA occurred in response to 200 J/mz UVR. This induction was partially blocked by treatment with PRL, indicating that DNA damage, the pyrimidine dimer in particular, played a role in uPA induction. In untransformed cultured corneal fibroblasts, the heparin-binding protein inhibitor, suramin, reduced basal uPA levels, UVR-induced uPA production and cell proliferation. Basic fibroblast growth factor, a heparin-binding growth factor known to be UVR-inducible in mesenchymal cells, stimulated uPA production and cell proliferation; however, anti-bFGF antibodies did not significantly decrease proliferation or basal uPA production. These findings suggested that basal levels of uPA secretion were modulated in response to heparin-binding growth factors and that these growth factors may also have mediated the effect of UVR on uPA levels.
[Posted on the website on 21 December 2000.
^Abbreviations: bFGF, basic fibroblast growth factor; DMEM, Dulbecco modified Eagle medium; ITS+ 1, insulin/transferrin/bovine serum albumin/sodium selenite/linoleic acid; MAPK, mitogen-activated protein kinase; PBS, phosphate-buffered saline; PRL, photoreactivating light; UVR, ultraviolet radiation.
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Wendy B. Green1, Paul G. McGuire', Katarzyna B. Miska2 and Donna F. Kusewitt*1
1Department of Cell Biology and Physiology, School of Medicine and 2Biology Department, University of New Mexico, Albuquerque, NM Received 11 May 2000; accepted 8 December 2000
*To whom correspondence should be addressed at: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, USA. Fax: 614-292-6473; e-mail: kusewitt.IC&osu.edu
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