Copper–GHK increases integrin expression and p63 positivity by keratinocytes
- 403 Downloads
Glycyl-l-histidyl-l-lysyl (GHK) possesses a high affinity for copper(II) ions, with which it spontaneously forms a complex (copper–GHK). It is well known that copper–GHK plays a physiological role in the process of wound healing and tissue repair by stimulating collagen synthesis in fibroblasts. This study was conducted to investigate the effects of copper–GHK on keratinocytes. Proliferative effects were analyzed and hematoxylin and eosin staining and immunohistochemistry were conducted to evaluate the effects of copper–GHK in skin equivalent (SE) models. In addition, western blotting was performed. In monolayer cultured keratinocytes, copper–GHK increased the proliferation of keratinocytes. When the SE models were evaluated, basal cells became cuboidal when copper–GHK was added. Immunohistochemical analysis revealed that copper–GHK increased proliferating cell nuclear antigen (PCNA) and p63 positivity. Furthermore, the expression of integrin α6 and β1 increased in SE models, and these results were confirmed by Western blotting. The results of this study indicate that treatment with copper–GHK may increase the proliferative potential of basal keratinocytes by modulating the expression of integrins, p63 and PCNA. In addition, increased levels of p63, a putative stem cell marker of the skin, suggests that copper–GHK promotes the survival of basal stem cells in the skin.
KeywordsPeptide Skin equivalents Integrin Keratinocytes Copper–GHK p63 Stem cells
This study was supported by a grant (A050432) from the Korea Health 21 R&D Project, Ministry of Health and Welfare.
- 10.Maquart FX, Bellon G, Chaqour B, Wegrowski J, Patt LM, Trachy RE, Monboisse JC, Chastang F, Birembaut P, Gillery P et al (1993) In vivo stimulation of connective tissue accumulation by the tripeptide-copper complex glycyl-l-histidyl-l-lysine-Cu2+ in rat experimental wounds. J Clin Invest 92:2368–2376. doi: 10.1172/JCI116842 PubMedCrossRefGoogle Scholar
- 12.Park KC, Choi HR, Cho HJ, Kim SK, Kwon SB, Kim DS, Cho YJ, Youn SW (2004) Effects of ascorbic acid on keratinocyte and epidermalization of skin. Ann Dermatol 16:45–51Google Scholar
- 18.Russell AJ, Fincher EF, Millman L, Smith R, Vela V, Waterman EA, Dey CN, Guide S, Weaver VM, Marinkovich MP (2003) Alpha 6 beta 4 integrin regulates keratinocyte chemotaxis through differential GTPase activation and antagonism of alpha 3 beta 1 integrin. J Cell Sci 116:3543–3556. doi: 10.1242/jcs.00663 PubMedCrossRefGoogle Scholar
- 19.Simeon A, Monier F, Emonard H, Gillery P, Birembaut P, Hornebeck W, Maquart FX (1999) Expression and activation of matrix metalloproteinases in wounds: modulation by the tripeptide-copper complex glycyl-l-histidyl-l-lysine-Cu2+. J Invest Dermatol 112:957–964. doi: 10.1046/j.1523-1747.1999.00606.x PubMedCrossRefGoogle Scholar
- 21.Sonnenberg A, Calafat J, Janssen H, Daams H, van der Raaij-Helmer LM, Falcioni R, Kennel SJ, Aplin JD, Baker J, Loizidou M et al (1991) Integrin alpha 6/beta 4 complex is located in hemidesmosomes, suggesting a major role in epidermal cell-basement membrane adhesion. J Cell Biol 113:907–917. doi: 10.1083/jcb.113.4.907 PubMedCrossRefGoogle Scholar
- 23.Tiberio R, Marconi A, Fila C, Fumelli C, Pignatti M, Krajewski S, Giannetti A, Reed JC, Pincelli C (2002) Keratinocytes enriched for stem cells are protected from anoikis via an integrin signaling pathway in a Bcl-2 dependent manner. FEBS Lett 524:139–144. doi: 10.1016/S0014-5793(02)03040-5 PubMedCrossRefGoogle Scholar