Increasing evidence shows persistent phenotypic alterations in fibroblasts from non-healing human chronic wounds, which may result in faulty extracellular matrix deposition and keratinocyte migration. We have previously shown that these cells are characterized by morphological changes, low proliferative potential and unresponsiveness to TGF-β1, and down regulated phosphorylation of Smad 2/3 and p42/44 MAPK from decreased expression of the TGF-β type II receptor.
To identify genes and proteins that may be differentially expressed in chronic wounds and their cultured fibroblasts.
Differential display analysis with 120 random primer sets was used in fibroblasts from human venous ulcers and acute wounds created on the ipsilateral thighs of the same patients. Positive differential results were confirmed by RT-PCR. Immunohistochemistry of cultured fibroblasts and tissues was used to determine the expression of differentially expressed proteins.
A total of 16 differentially expressed genes were identified and cloned. The only candidate gene that was differentially expressed in all patients and confirmed by repeated differential display testing and RT-PCR was βig-h3, a TGF-β-induced gene involved in cell adhesion, migration, and proliferation. Decreased expression of βig-h3 in chronic wounds and their fibroblasts was further confirmed by Western blot and immunostaining.
These findings point to βig-h3 as an important gene characterizing the abnormal phenotype of chronic wound fibroblasts. Corrective measures to increase the expression of this protein might have therapeutic potential.
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- Wound healing and its impairment in the diabetic foot.Lancet. 2005; 366: 1736-1743
- Fibroblasts from chronic wounds show altered TGF-beta-signaling and decreased TGF-beta type II receptor expression.J Cell Physiol. 2003; 195: 331-336
- Defective extracellular matrix reorganization by chronic wound fibroblasts is associated with alterations in TIMP-1, TIMP-2, and MMP-2 activity.J Invest Dermatol. 2000; 115: 225-233
- Proliferation and mitogenic response to PDGF-BB of fibroblasts isolated from chronic venous leg ulcers is ulcer-age dependent.J Invest Dermatol. 1999; 112: 463-469
- Dermal fibroblasts from venous ulcers are unresponsive to the action of transforming growth factor-beta 1.J Dermatol Sci. 1997; 16: 59-66
- Decreased levels of alpha 1(I) procollagen mRNA in dermal fibroblasts grown on fibrin gels and in response to fibrinopeptide B.J Cell Physiol. 1995; 162: 9-14
- Annealing control primer system for identification of differentially expressed genes on agarose gels.Biotechniques. 2004; 36 (428, 430 passim): 424-426
- Human beta-defensin-2 expression is increased in chronic wounds.Wound Repair Regen. 2004; 12: 439-443
- Full-thickness wounding of the mouse tail as a model for delayed wound healing: accelerated wound closure in Smad3 knock-out mice.Wound Repair Regen. 2004; 12: 320-326
- Effect of basement membrane entactin on epidermal cell attachment and growth.J Invest Dermatol. 1987; 88: 55-59
- Annealing control primer system for improving specificity of PCR amplification.Biotechniques. 2003; 35: 1180-1184
- Identification of differentially regulated genes in bovine blastocysts using an annealing control primer system.Mol Reprod Dev. 2004; 69: 43-51
- Methods in molecular biology.in: Meade J.D. Liang P. Pardee A.B. Differential display methods and protocols. 2nd ed. Humana Press Inc., Totowa2005
- The protein components of the 12-nanometer microfibrils of elastic and nonelastic tissues.J Biol Chem. 1989; 264: 4590-4598
- Beta ig-h3: a transforming growth factor-beta-responsive gene encoding a secreted protein that inhibits cell attachment in vitro and suppresses the growth of CHO cells in nude mice.DNA Cell Biol. 1994; 13: 571-584
- cDNA cloning and sequence analysis of beta ig-h3, a novel gene induced in a human adenocarcinoma cell line after treatment with transforming growth factor-beta.DNA Cell Biol. 1992; 11: 511-522
- Characterization of a cartilage-derived 66-kDa protein (RGD-CAP/beta ig-h3) that binds to collagen.Biochim Biophys Acta. 1997; 1355: 303-314
- Immunohistochemical and ultrastructural localization of MP78/70 (betaig-h3) in extracellular matrix of developing and mature bovine tissues.J Histochem Cytochem. 1997; 45: 1683-1696
- Expression of betaig-h3 by human bronchial smooth muscle cells: localization to the extracellular matrix and nucleus.Am J Respir Cell Mol Biol. 2000; 22: 352-359
- Betaig-h3, a novel secretory protein inducible by transforming growth factor-beta, is present in normal skin and promotes the adhesion and spreading of dermal fibroblasts in vitro.J Invest Dermatol. 1995; 104: 844-849
- Beta-ig: molecular cloning and in situ hybridization in corneal tissues.Invest Ophthalmol Vis Sci. 1997; 38: 893-900
- Accumulation of betaig-h3 gene product in corneas with granular dystrophy.Am J Pathol. 1998; 152: 743-748
- Kerato-epithelin mutations in four 5q31-linked corneal dystrophies.Nat Genet. 1997; 15: 247-251
- Transforming growth factor-beta up-regulates the beta 5 integrin subunit expression via Sp1 and Smad signaling.J Biol Chem. 2000; 275: 36400-36406
- TGF-beta1-stimulated osteoblasts require intracellular calcium signaling for enhanced alpha5 integrin expression.Ann NY Acad Sci. 2002; 961: 178-182
- Rapid up-regulation of alpha4 integrin-mediated leukocyte adhesion by transforming growth factor-beta1.Mol Biol Cell. 2003; 14: 54-66
- Identification of motifs in the fasciclin domains of the transforming growth factor-beta-induced matrix protein betaig-h3 that interact with the alphavbeta5 integrin.J Biol Chem. 2002; 277: 46159-46165
- TGF-beta1 enhances betaig-h3-mediated keratinocyte cell migration through the alpha3beta1 integrin and PI3K.J Cell Biochem. 2004; 92: 770-780
- Identification of the alphavbeta3 integrin-interacting motif of betaig-h3 and its anti-angiogenic effect.J Biol Chem. 2003; 278: 25902-25909
- Mice lacking Smad3 show accelerated wound healing and an impaired local inflammatory response.Nat Cell Biol. 1999; 1: 260-266
Published online: July 12, 2010
Accepted: October 17, 2007
Received in revised form: October 13, 2007
Received: February 25, 2007
© 2007 Published by Elsevier Inc.