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Archives of Dermatological Research

, Volume 300, Issue 6, pp 303–310 | Cite as

SERPINE1 (PAI-1) is deposited into keratinocyte migration “trails” and required for optimal monolayer wound repair

  • Kirwin M. Providence
  • Stephen P. Higgins
  • Andrew Mullen
  • Ashley Battista
  • Rohan Samarakoon
  • Craig E. Higgins
  • Cynthia E. Wilkins-Port
  • Paul J. Higgins
Original Paper

Abstract

Cutaneous tissue injury, both in vivo and in vitro, initiates activation of a “wound repair” transcriptional program. One such highly induced gene encodes plasminogen activator inhibitor type-1 (PAI-1, SERPINE1). PAI-1-GFP, expressed as a fusion protein under inducible control of +800 bp of the wound-activated PAI-1 promoter, prominantly “marked” keratinocyte migration trails during the real-time of monolayer scrape-injury repair. Addition of active recombinant PAI-1 to wounded wild-type keratinocyte monolayers as well as to PAI-1−/− MEFs and PAI-1−/− keratinocytes significantly stimulated directional motility above basal levels in all cell types. PAI-1 expression knockdown or antibody-mediated functional inhibition, in contrast, effectively attenuated injury repair. The defect in wound-associated migratory activity as a consequence of antisense-mediated PAI-1 down-regulation was effectively reversed by addition of recombinant PAI-1 immediately after scrape injury. One possible mechanism underlying the PAI-1-dependent motile response may involve fine control of the keratinocyte substrate detachment/re-attachment process. Exogenous PAI-1 significantly enhanced keratinocyte spread cell “footprint” area while PAI-1 neutralizing antibodies, but not control non-immune IgG, effectively inhibited spreading with apoptotic hallmarks evident within 24 h. Importantly, PAI-1 not only stimulated keratinocyte adhesion and wound-initiated planar migration but also rescued keratinocytes from plasminogen-induced substrate detachment/anoikis. The early transcriptional response of the PAI-1 gene to monolayer trauma and its prominence in the injury repair genetic signature are consistent with its function as both a survival factor and regulator of the time course of epithelial migration as part of the cutaneous injury response program.

Keywords

PAI-1 SERPINE1 Wound healing Keratinocytes Gene targeting Migration 

Notes

Acknowledgments

Supported by NIH grant GM57242.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Kirwin M. Providence
    • 1
  • Stephen P. Higgins
    • 2
  • Andrew Mullen
    • 1
  • Ashley Battista
    • 1
  • Rohan Samarakoon
    • 2
  • Craig E. Higgins
    • 2
  • Cynthia E. Wilkins-Port
    • 2
  • Paul J. Higgins
    • 2
  1. 1.Department of BiologyIthaca CollegeIthacaUSA
  2. 2.Center for Cell Biology and Cancer ResearchAlbany Medical College (Mail Code 165)AlbanyUSA

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