Archives of Dermatological Research

, Volume 304, Issue 7, pp 533–540 | Cite as

Protein profiling of keloidal scar tissue

Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

Clinically, keloids are defined as scars that invade adjacent healthy tissue and are caused by tissue injury; however, the clear distinction relating keloids to wound healing or to cancer remains elusive. This study profiled protein extracts from the regions of keloid tissues to determine the activities within these different zones, and to help underpin the activities apparent within different regions of the disease. Two-dimensional gel electrophoresis, liquid chromatography mass spectrometer (LCMS) and a Mascot online database search were employed for comparative proteomic analysis between four sites in keloidal scars (KS). Out of 400 spots identified in all gels, 21 were unique to given KS sites. These were identified using LCMS and the results showed the presence of mitochondrial and structural proteins in the margin, while the top contained keratin II. Heat shock protein was the only protein present in the internal normal control and margin, while the external normal contain keratin II and the voltage-dependent anion-selective channel protein, annexin A6, plus glial fibrillary acidic protein. This work is novel since it has identified differentially expressed proteins across the tested keloidal scar sites. The presence of mitochondrial-associated proteins at the margins suggests that this is the most active part of the scar.

Keywords

Keloid protein profile Two-dimensional gel electrophoresis Liquid chromatography mass spectrometer 
This is a preview of subscription content, log in to check access.

References

  1. 1.
    Alonso PE, Rioja LF, Pera C (2008) Keloids: a viral hypothesis. Med Hypotheses 70(1):156–166PubMedCrossRefGoogle Scholar
  2. 2.
    Billack B, Heck DE, Mariano TM, Gardner CR, Sur R, Laskin DL, Laskin JD (2002) Induction of cyclooxygenase-2 by heat shock protein 60 in macrophages and endothelial cells. Am J Physiol Cell Physiol 283(4):C1267–C1277PubMedGoogle Scholar
  3. 3.
    Brody GS (1993) Biological creep. Plast Reconstr Surg 92(6):1202–1203PubMedGoogle Scholar
  4. 4.
    Calderon M, Lawrence WT, Banes AJ (1996) Increased proliferation in keloid fibroblasts wounded in vitro. J Surg Res 61(2):343–347PubMedCrossRefGoogle Scholar
  5. 5.
    Chuang NN, Huang CC (2007) Interaction of integrin beta1 with cytokeratin 1 in neuroblastoma NMB7 cells. Biochem Soc Trans 35(Pt 5):1292–1294PubMedGoogle Scholar
  6. 6.
    Diao JS, Xia WS, Yi CG, Wang YM, Li B, Xia W, Liu B, Guo SZ, Sun XD (2011) Trichostatin A inhibits collagen synthesis and induces apoptosis in keloid fibroblasts. Arch Dermatol Res 303(8):573–580PubMedCrossRefGoogle Scholar
  7. 7.
    Diegelmann RF, Cohen IK, McCoy BJ (1979) Growth kinetics and collagen synthesis of normal skin, normal scar and keloid fibroblasts in vitro. J Cell Physiol 98(2):341–346PubMedCrossRefGoogle Scholar
  8. 8.
    Dienus K, Bayat A, Gilmore BF, Seifert O (2010) Increased expression of fibroblast activation protein-alpha in keloid fibroblasts: implications for development of a novel treatment option. Arch Dermatol Res 302(10):725–731PubMedCrossRefGoogle Scholar
  9. 9.
    Doherty GJ, McMahon HT (2008) Mediation, modulation, and consequences of membrane-cytoskeleton interactions. Annu Rev Biophys 37:65–95PubMedCrossRefGoogle Scholar
  10. 10.
    Fong EP, Bay BH (2002) Keloids: the sebum hypothesis revisited. Med Hypotheses 58:264PubMedCrossRefGoogle Scholar
  11. 11.
    Fujiwara M, Muragaki Y, Ooshima A (2005) Upregulation of transforming growth factor-beta1 and vascular endothelial growth factor in cultured keloid fibroblasts: relevance to angiogenic activity. Arch Dermatol Res 297(4):161–169PubMedCrossRefGoogle Scholar
  12. 12.
    Haisa M, Okochi H, Grotendorst GR (1994) Elevated levels of PDGF alpha receptors in keloid fibroblasts contribute to an enhanced response to PDGF. J Invest Dermatol 103(4):560–563PubMedCrossRefGoogle Scholar
  13. 13.
    Hiller S, Abramson J, Mannella C, Wagner G, Zeth K (2010) The 3D structures of VDAC represent a native conformation. Trends Biochem Sci 35(9):514–521PubMedCrossRefGoogle Scholar
  14. 14.
    Jaattela M, Wissing D (1992) Emerging role of heat shock proteins in biology and medicine. Ann Med 24(4):249–258PubMedCrossRefGoogle Scholar
  15. 15.
    Katsumoto T, Mitsushima A, Kurimura T (1990) The role of the vimentin intermediate filaments in rat 3Y1 cells elucidated by immunoelectron microscopy and computer-graphic reconstruction. Biol Cell 68(2):139–146PubMedCrossRefGoogle Scholar
  16. 16.
    Khoo YT, Ong CT, Mukhopadhyay A, Han HC, Do DV, Lim IJ, Phan TT (2006) Upregulation of secretory connective tissue growth factor (CTGF) in keratinocyte-fibroblast coculture contributes to keloid pathogenesis. J Cell Physiol 208(2):336–343PubMedCrossRefGoogle Scholar
  17. 17.
    Knowles JR (1980) Enzyme-catalyzed phosphoryl transfer reactions. Annu Rev Biochem 49:877–919PubMedCrossRefGoogle Scholar
  18. 18.
    Leube RE, Bader BL, Bosch FX, Zimbelmann R, Achtstaetter T, Franke WW (1988) Molecular characterization and expression of the stratification-related cytokeratins 4 and 15. J Cell Biol 106(4):1249–1261PubMedCrossRefGoogle Scholar
  19. 19.
    Lopes LB, Flynn C, Komalavilas P, Panitch A, Brophy CM, Seal BL (2009) Inhibition of HSP27 phosphorylation by a cell-permeant MAPKAP Kinase 2 inhibitor. Biochem Biophys Res Commun 382(3):535–539PubMedCrossRefGoogle Scholar
  20. 20.
    Marneros AG, Krieg T (2004) Keloids—clinical diagnosis, pathogenesis, and treatment options. J Dtsch Dermatol Ges 2(11):905–913PubMedCrossRefGoogle Scholar
  21. 21.
    Morris SD (2002) Heat shock proteins and the skin. Clin Exp Dermatol 27(3):220–224PubMedCrossRefGoogle Scholar
  22. 22.
    Mukhopadhyay A, Chan SY, Lim IJ, Phillips DJ, Phan TT (2007) The role of the activin system in keloid pathogenesis. Am J Physiol Cell Physiol 292(4):C1331–C1338PubMedCrossRefGoogle Scholar
  23. 23.
    Naciff JM, Behbehani MM, Kaetzel MA, Dedman JR (1996) Annexin VI modulates Ca2+ and K+ conductances of spinal cord and dorsal root ganglion neurons. Am J Physiol 271(6 Pt 1):C2004–C2015PubMedGoogle Scholar
  24. 24.
    Niessen FB, Spauwen PH, Schalkwijk J, Kon M (1999) On the nature of hypertrophic scars and keloids: a review. Plast Reconstr Surg 104(5):1435–1458PubMedCrossRefGoogle Scholar
  25. 25.
    Njemini R, Lambert M, Demanet C, Mets T (2003) Elevated serum heat-shock protein 70 levels in patients with acute infection: use of an optimized enzyme-linked immunosorbent assay. Scand J Immunol 58(6):664–669PubMedCrossRefGoogle Scholar
  26. 26.
    O’Reilly AM, Currie RW, Clarke DB (2010) HspB1 (Hsp 27) expression and neuroprotection in the retina. Mol Neurobiol 42(2):124–132PubMedCrossRefGoogle Scholar
  27. 27.
    Ong CT, Khoo YT, Mukhopadhyay A, Masilamani J, Do DV, Lim IJ, Phan TT (2010) Comparative proteomic analysis between normal skin and keloid scar. Br J Dermatol 162(6):1302–1315PubMedCrossRefGoogle Scholar
  28. 28.
    Ong CT, Khoo YT, Tan EK, Mukhopadhyay A, Do DV, Han HC, Lim IJ, Phan TT (2007) Epithelial-mesenchymal interactions in keloid pathogenesis modulate vascular endothelial growth factor expression and secretion. J Pathol 211(1):95–108PubMedCrossRefGoogle Scholar
  29. 29.
    Pappin DJ, Hojrup P, Bleasby AJ (1993) Rapid identification of proteins by peptide-mass fingerprinting. Curr Biol 3(6):327–332PubMedCrossRefGoogle Scholar
  30. 30.
    Prakken BJ, Roord S, Ronaghy A, Wauben M, Albani S, van Eden W (2003) Heat shock protein 60 and adjuvant arthritis: a model for T cell regulation in human arthritis. Springer Semin Immunopathol 25(1):47–63PubMedCrossRefGoogle Scholar
  31. 31.
    Sheehan D, Meade G, Foley VM, Dowd CA (2001) Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochem J 360(Pt 1):1–16PubMedCrossRefGoogle Scholar
  32. 32.
    Shih B, Bayat A (2010) Genetics of keloid scarring. Arch Dermatol Res 302(5):319–339PubMedCrossRefGoogle Scholar
  33. 33.
    Smith FJ, Maingi C, Covello SP, Higgins C, Schmidt M, Lane EB, Uitto J, Leigh IM, McLean WH (1998) Genomic organization and fine mapping of the keratin 2e gene (KRT2E): K2e V1 domain polymorphism and novel mutations in ichthyosis bullosa of Siemens. J Invest Dermatol 111(5):817–821PubMedCrossRefGoogle Scholar
  34. 34.
    Su CW, Alizadeh K, Boddie A, Lee RC (1998) The problem scar. Clin Plast Surg 25(3):451–465PubMedGoogle Scholar
  35. 35.
    Switzer RC 3rd, Merril CR, Shifrin S (1979) A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal Biochem 98(1):231–237PubMedCrossRefGoogle Scholar
  36. 36.
    Tanaka A, Hatoko M, Tada H, Iioka H, Niitsuma K, Miyagawa S (2004) Expression of p53 family in scars. J Dermatol Sci 34(1):17–24PubMedCrossRefGoogle Scholar
  37. 37.
    Tang B, Zhu B, Liang Y, Bi L, Hu Z, Chen B, Zhang K, Zhu J (2011) Asiaticoside suppresses collagen expression and TGF-beta/Smad signaling through inducing Smad7 and inhibiting TGF-betaRI and TGF-betaRII in keloid fibroblasts. Arch Dermatol Res 303(8):563–572PubMedCrossRefGoogle Scholar
  38. 38.
    Tardy M, Fages C, Le Prince G, Rolland B, Nunez J (1990) Regulation of the glial fibrillary acidic protein (GFAP) and of its encoding mRNA in the developing brain and in cultured astrocytes. Adv Exp Med Biol 265:41–52PubMedGoogle Scholar
  39. 39.
    Totan S, Echo A, Yuksel E (2011) Heat shock proteins modulate keloid formation. Eplasty 11:e21PubMedGoogle Scholar
  40. 40.
    Wallimann T, Wyss M, Brdiczka D, Nicolay K, Eppenberger HM (1992) Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: the ‘phosphocreatine circuit’ for cellular energy homeostasis. Biochem J 281(Pt 1):21–40PubMedGoogle Scholar
  41. 41.
    Wang JF, Olson ME, Reno CR, Kulyk W, Wright JB, Hart DA (2000) Molecular and cell biology of skin wound healing in a pig model. Connect Tissue Res 41(3):195–211PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  1. 1.Quantitative Molecular Medicine Group, Manchester Interdisciplinary Biocentre, School of Cancer and Enabling SciencesUniversity of ManchesterManchesterUK

Personalised recommendations