Skip to main content

Advertisement

SpringerLink
Log in

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

  • Original Paper
  • Published:
Archives of Dermatological Research Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Allen RR, Qi L, Higgins PJ (2005) Upstream stimulatory factor regulates E box-dependent PAI-1 transcription in human epidermal keratinocytes. J Cell Physiol 203:156–165

    Article  PubMed  CAS  Google Scholar 

  2. Andreasen PA, Egelund R, Peterson HH (2000) The plasminogen activation system in tumor growth, invasion, and metastasis. Cell Mol Life Sci 57:25–40

    Article  PubMed  CAS  Google Scholar 

  3. Bajou K, Masson V, Gerard RD, Schmitt PM, Albert V, Praus M, Lund LR, Frandsen TL, Brunner N, Dano K, Fusenig NE, Weidle U, Carmeliet G, Loskutoff D, Collen D, Carmeliet P, Foidart JM, Noel A (2001) The plasminogen activator inhibitor PAI-1 controls in vivo tumor vascularization by interaction with proteases, not vitronectin. Implications for antiangiogenic strategies. J Cell Biol 152:777–784

    Article  PubMed  CAS  Google Scholar 

  4. Bretland AJ, Lawry J, Sharrard RM (2001) A study of death by anoikis in cultured epithelial cells. Cell Prolif 34:199–210

    Article  PubMed  CAS  Google Scholar 

  5. Brooks TD, Slomp J, Quax PHA, De Bart ACW, Spencer MT, Verheijen JH, Charlton PA (2001) Antibodies to PAI-1 alter the invasive and migratory properties of human tumour cells in vitro. Clin Exp Metastasis 18:445–453

    Article  Google Scholar 

  6. Chapman HA, Wei Y (2001) Protease crosstalk with integrins: the urokinase receptor paradigm. Thromb Haemost 86:124–129

    PubMed  CAS  Google Scholar 

  7. Chazaud B, Ricoux R, Christov C, Plonquet A, Gherardi RK, Barlovatz-Meimon G (2002) Promigratory effect of plasminogen activator inhibitor-1 on invasive breast cancer populations. Am J Pathol 160:237–246

    PubMed  CAS  Google Scholar 

  8. Czekay R-P, Aertgeerts K, Curriden SA, Loskutoff DJ (2003) Plasminogen activator inhibitor-1 detaches cells from extracellular matrices by inactivating Integrins. J Cell Biol 160:781–791

    Article  PubMed  CAS  Google Scholar 

  9. Degryse B, Neels JG, Czekay R-P, Aertgeerts K, Kamikubo T, Loskutoff DJ (2004) The low density lipoprotein receptor-related protein is a motogenic receptor for plasminogen activator inhibitor-1. J Biol Chem 279:22595–22604

    Article  PubMed  CAS  Google Scholar 

  10. Deng G, Curriden SA, Hu G, Czekay R-P, Loskutoff DJ (2001) Plasminogen activator inhibitor-1 regulates cell adhesion by binding to the somatomedin B domain of vitronectin. J Cell Physiol 189:23–33

    Article  PubMed  CAS  Google Scholar 

  11. Dieckgraefe BK, Weems DM, Santoro SA, Alpers DH (1997) p38 MAP kinase pathways are mediators of intestinal epithelial wound-induced signal transduction. Biochem Biophys Res Commun 233:23–33

    Article  Google Scholar 

  12. Fitsialos G, Chassot AA, Turchi L, Dayem MA, LeBrigand K, Moreilhon C, Meneguzzi G, Busca R, Mari B, Barbry P, Ponzio G (2007) Transcriptional signature of epidermal keratinocytes subjected to in vitro scratch wounding reveals selective roles for ERK1/2, p38, and phosphatidylinositol 3-kinase signaling pathways. J Biol Chem 282:15090–15102

    Article  PubMed  CAS  Google Scholar 

  13. Geer DJ, Andreadis ST (2003) A novel role of fibrin in epidermal healing: plasminogen-mediated migration and selective detachment of differentiated keratinocytes. J Invest Dermatol 121:1210–1216

    Article  PubMed  CAS  Google Scholar 

  14. Gutierrez LS, Schulman A, Brito-Robinson T, Noria F, Ploplis VA, Castellino FJ (2000) Tumor development is retarded in mice lacking the gene for urokinase-type plasminogen activator or its inhibitor, plasminogen activator inhibitor-1. Cancer Res 60:5839–5847

    PubMed  CAS  Google Scholar 

  15. Higgins PJ, Ryan MP, Jelley DM (1997) p52PAI-1 gene expression in butyrate-induced flat revertants of v-ras-transformed rat kidney cells: mechanism of induction and involvement in the morphologic response. Biochem J 321:431–437

    PubMed  CAS  Google Scholar 

  16. Higgins PJ, Slack JK, Diegelmann RF, Staiano-Coico L (1999) Differential regulation of PAI-1 gene expression in human fibroblasts predisposed to a fibrotic phenotype. Exp Cell Res 248:634–642

    Article  PubMed  CAS  Google Scholar 

  17. Iyer VR, Eisen MB, Ross DT, Schuler G, Moore T, Lee JC, Trent JM, Staudt LM, Hudson J, Boguski MS, Lashkari D, Shalon D, Botstein D, Brown PO (1999) The transcriptional program in the response of human fibroblasts to serum. Science 283:83–87

    Article  PubMed  CAS  Google Scholar 

  18. Jensen PJ, Lavker RM (1996) Modulation of the plasminogen activator cascade during enhanced epidermal proliferation in vivo. Cell Growth Differ 7:1793–1804

    PubMed  CAS  Google Scholar 

  19. Kortlever RM, Higgins PJ, Bernards R (2006) Plasminogen activator inhibitor-1 is a critical downstream target of p53 in the induction of replicative senescence. Nat Cell Biol 8:877–884

    Article  PubMed  CAS  Google Scholar 

  20. Kutz SM, Nickey S, White LA, Higgins PJ (1997) Induced PAI-1 mRNA expression and targeted protein accumulation are early G1 events in serum-stimulated rat kidney cells. J Cell Physiol 170:8–18

    Article  PubMed  CAS  Google Scholar 

  21. Maquerlot F, Galiacy S, Malo M, Guignabert C, Lawrence DA, d’Ortho MP, Barlovatz-Meimon G (2006) Dual role for plasminogen activator inhibitor type 1 as a soluble and as matricellular regulator of epithelial alveolar cell healing. Am J Pathol 169:1624–1632

    Article  PubMed  CAS  Google Scholar 

  22. Martin P (1997) Wound healing—aiming for perfect skin regeneration. Science 176:75–81

    Article  Google Scholar 

  23. Mazzalupo S, Wawersik MJ, Coulombe PA (2002) An ex vivo assay to assess the potential of skin keratinocytes for wound epithelialization. J Invest Dermatol 118:866–870

    Article  PubMed  CAS  Google Scholar 

  24. Palmeri D, Lee JW, Juliano RL, Church FC (2002) Plasminogen activator inhibitor-1 and -3 increase cell adhesion and motility in MDA-MB-435 breast cancer cells. J Biol Chem 277:40950–40957

    Article  Google Scholar 

  25. Pawar S, Kartha S, Tobeck FG (1995) Differential gene expression in migrating renal epithelial cells after wounding. J Cell Physiol 165:556–565

    Article  PubMed  CAS  Google Scholar 

  26. Planus E, Barlovatz-Meimon G, Rogers RA, Bonavaud S, Ingber DF, Wang N (1997) Binding of urokinase to plasminogen activator inhibitor type-1 mediates cell adhesion and spreading. J Cell Sci 110:1091–1098

    PubMed  CAS  Google Scholar 

  27. Providence KM, Higgins PJ (2004) PAI-1 expression is required for epithelial cell migration in two distinct phases of in vitro wound repair. J Cell Physiol 200:297–308

    Article  PubMed  CAS  Google Scholar 

  28. Providence KM, Staiano-Coico L, Higgins PJ (2003) A quantifiable in vitro model to assess effects of PAI-1 gene targeting on epithelial cell motility. Methods Mol Med 78:293–303

    PubMed  CAS  Google Scholar 

  29. Providence KM, White LA, Tang J, Gonclaves J, Staiano-Coico L, Higgins PJ (2002) Epithelial monolayer wounding stimulates binding of USF-1 to an E-box motif in the plasminogen activator inhibitor type 1 gene. J Cell Sci 115:3767–3777

    Article  PubMed  CAS  Google Scholar 

  30. Qi L, Allen RR, Lu Q, Higgins CE, Garone R, Staiano-Coico L, Higgins PJ (2006) PAI-1 transcriptional regulation during the G0→G1 transition in human epidermal keratinocytes. J Cell Biochem 99:495–507

    Article  PubMed  CAS  Google Scholar 

  31. Qi L, Higgins SP, Lu Q, Samarakoon R, Wilkins-Port CE, Ye Q, Higgins CE, Staiano-Coico L, Higgins PJ (2007) SERPINE1 (PAI-1) is a prominent member of the early G0→G1 transition “wound repair” transcriptome in p53 mutant human keratinocytes. J Invest Dermatol 128:749–753

    Article  PubMed  CAS  Google Scholar 

  32. Reinartz J, Link J, Todd RF, Kramer MD (1994) The receptor for urokinase-type plasminogen activator of a human keratinocyte line (HaCaT). Exp Cell Res 214:486–498

    Article  PubMed  CAS  Google Scholar 

  33. Romer MU, Kirkebjerg Due A, Knud Larsen J, Hofland KF, Christensen IJ, Buhl-Jensen P, Almholt K, Lerberg Nielsen O, Brunner N, Lademann U (2005) Indication of a role of plasminogen activator inhibitor type 1 in protecting murine fibrosarcoma cells against apoptosis. Thromb Haemost 94:859–866

    PubMed  Google Scholar 

  34. Romer J, Lund LR, Kriksen J, Ralkiaer E, Zeheb R, Gelehrter TD, Dano K, Kristensen P (1991) Differential expressin of urokinase-type plasminogen activator and its type-1 inhibitor during healing of mouse skin wounds. J Invest Dermatol 102:519–522

    Article  Google Scholar 

  35. Rossignol P, Ho-Tin-Noe B, Vranckx R, Bouton MC, Meilhac O, Lijnen HR, Guillin MC, Michel JB, Angles-Cano E (2004) Protease nexin-1 inhibits plasminogen activation-induced apoptosis of adherent cells. J Biol Chem 279:10346–10356

    Article  PubMed  CAS  Google Scholar 

  36. Seebacher T, Manske M, Zoller J, Crabb J, Bade EG (1992) The EGF-inducible protein EIP-1 of migrating normal and malignant rat liver epithelial cells is identical to plasminogen activator inhibitor 1 and is a component of the ECM migration tracks. Exp Cell Res 203:504–507

    Article  PubMed  CAS  Google Scholar 

  37. Stefansson S, Lawrence DA (2003) Old dogs and new tricks: proteases, inhibitors, and cell migration. Sci STKE 2003(189):24

    Article  Google Scholar 

  38. Tuan TL, Zhu JY, Sun B, Nichter LS, Nimni ME, Laug WE (1996) Elevated levels of plasminogen activator inhibitor-1 may account for the altered fibrinolysis by keloid fibroblasts. J Invest Dermatol 106:1007–1011

    Article  PubMed  CAS  Google Scholar 

  39. Turchi L, Chassot AA, Rezzonico R, Yeow K, Loubat A, Ferrua B, Lenegrate G, Ortonne JP, Ponzio G (2002) Dynamic characterization of the molecular events during in vitro epidermal wound healing. J Invest Dermatol 119:56–63

    Article  PubMed  CAS  Google Scholar 

  40. Wang Z, Sosne G, Kurpakus-Wheater M (2005) Plasminogen activator inhibitor-1 (PAI-1) stimulates human corneal epithelial cell adhesion and migration in vitro. Exp Eye Res 80:1–8

    Article  PubMed  CAS  Google Scholar 

  41. Wawersik MJ, Mazzalupo S, Nguyen D, Coulombe PA (2001) Increased levels of keratin 16 alter epithelialization potential of mouse skin keratinocytes in vivo and ex vivo. Mol Biol Cell 12:3439–3450

    PubMed  CAS  Google Scholar 

  42. Wilkins-Port CE, Higgins PJ (2007) Regulation of extracellular matrix remodeling following TGF-β1/EGF-stimulated epithelial-mesenchymal transition in human pre-malignant keratinocytes. Cells Tissues Organs 185:116–122

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Supported by NIH grant GM57242.

Author information

Authors and Affiliations

  1. Department of Biology, Ithaca College, Ithaca, NY, 14850, USA

    Kirwin M. Providence, Andrew Mullen & Ashley Battista

  2. Center for Cell Biology and Cancer Research, Albany Medical College (Mail Code 165), 47 New Scotland Avenue, Albany, NY, 12208, USA

    Stephen P. Higgins, Rohan Samarakoon, Craig E. Higgins, Cynthia E. Wilkins-Port & Paul J. Higgins

Authors
  1. Kirwin M. Providence
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stephen P. Higgins
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andrew Mullen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ashley Battista
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rohan Samarakoon
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Craig E. Higgins
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Cynthia E. Wilkins-Port
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Paul J. Higgins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Paul J. Higgins.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Providence, K.M., Higgins, S.P., Mullen, A. et al. SERPINE1 (PAI-1) is deposited into keratinocyte migration “trails” and required for optimal monolayer wound repair. Arch Dermatol Res 300, 303–310 (2008). https://doi.org/10.1007/s00403-008-0845-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00403-008-0845-2

Keywords

Search

Navigation