Research Article| Volume 71, ISSUE 2, P115-121, August 2013

Download started.


Expressed in high metastatic cells (Ehm2) is a positive regulator of keratinocyte adhesion and motility: The implication for wound healing



      Multiple factors have been shown to delay dermal wound healing. These resultant wounds pose a significant problem in terms of morbidity and healthcare spend. Recently, an increasing volume of research has focused on the molecular perturbations underlying non-healing wounds.


      This study investigates the effect of a novel cancer promoter, Ehm2, in wound healing. Ehm2 belongs to the FERM family of proteins, known to be involved in membrane–cytoskeletal interactions, and has been shown to promote cancer metastasis in melanoma, prostate cancer and breast cancer.


      Ehm2 mRNA levels were analysed using qRT-PCR, standardised to GAPDH, from either acute or chronic wounds, and normal skin. IHC analysis was also undertaken from wound edge biopsies. An anti-Ehm2 transgene was created and transfected into the HaCaT cell line. The effect of Ehm2 knockdown on migration, adhesion, growth, cell cycle progression and apoptosis was analysed using standard laboratory methods. Western Blot analysis was used to investigate potential downstream protein interactions.


      Ehm2 is expressed nearly three times higher in acute wound tissues, compared to chronic wound tissues. Increased Ehm2 expression is found in wounds undergoing healing, especially at the leading wound edge. In vitro, Ehm2 knockdown reduces cellular adhesion, migration and motility, without affecting growth, cell cycle and apoptosis. Finally, Ehm2 knockdown results in reduced NWasp protein expression.


      These results suggest Ehm2 may be an important player in the wound healing process, and show that Ehm2 knockdown downregulates the expression of NWasp, through which it may have its effect on cellular migration.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of Dermatological Science
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Velnar T.
        • Bailey T.
        • Smrkolj V.
        The wound healing process: an overview of the cellular and molecular mechanisms.
        J Int Med Res. 2009; 37: 1528-1542
        • Harding K.G.
        • Morris H.L.
        • Patel G.K.
        Science, medicine, and the future – healing chronic wounds.
        Br Med J. 2002; 324: 160-163
        • Medina A.
        • Scott P.G.
        • Ghahary A.
        • Tredget E.E.
        Pathophysiology of chronic nonhealing wounds.
        J Burn Care Rehabil. 2005; 26: 306-319
        • Sanders A.J.
        • Jiang D.G.
        • Jiang W.G.
        • Harding K.G.
        • Patel G.K.
        Activated leukocyte cell adhesion molecule impacts on clinical wound healing and inhibits HaCaT migration.
        Int Wound J. 2011; 8: 500-507
        • Jiang W.G.
        • Ye L.
        • Patel G.
        • Harding K.G.
        Expression of WAVEs, the WASP (Wiskott–Aldrich syndrome protein) family of verprolin homologous proteins in human wound tissues and the biological influence on human keratinocytes.
        Wound Repair Regen. 2010; 18: 594-604
        • Stojadinovic O.
        • Pastar I.
        • Vukelic S.
        • Mahoney M.G.
        • Brennan D.
        • Krzyzanowska A.
        • et al.
        Deregulation of keratinocyte differentiation and activation: a hallmark of venous ulcers.
        J Cell Mol Med. 2008; 12: 2675-2690
        • Kirsner R.S.
        • Charles C.A.
        • Tomic-Canic M.
        • Vincek V.
        • Nassiri M.
        • Stojadinovic O.
        • et al.
        A gene signature of nonhealing venous ulcers: potential diagnostic markers.
        J Am Acad Dermatol. 2008; 59: 758-771
        • Stojadinovic O.
        • Brem H.
        • Vouthounis C.
        • Lee B.
        • Fallon J.
        • Stallcup M.
        • et al.
        Molecular pathogenesis of chronic wounds: the role of beta-catenin and c-myc in the inhibition of epithelialization and wound healing.
        Am J Pathol. 2005; 167: 59-69
        • Dvorak H.F.
        Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing.
        N Engl J Med. 1986; 315: 1650-1659
        • Hashimoto Y.
        • Shindo-Okada N.
        • Tani M.
        • Takeuchi K.
        • Toma H.
        • Yokota J.
        Identification of genes differentially expressed in association with metastatic potential of K-1735 murine melanoma by messenger RNA differential display.
        Cancer Res. 1996; 56: 5266-5271
        • Shimizu K.
        • Nagamachi Y.
        • Tani M.
        • Kimura K.
        • Shiroishi T.
        • Wakana S.
        • et al.
        Molecular cloning of a novel NF2/ERM/4.1 superfamily gene, ehm2, that is expressed in high-metastatic K1735 murine melanoma cells.
        Genomics. 2000; 65: 113-120
        • Sun C.X.
        • Robb V.A.
        • Gutmann D.H.
        Protein 4.1 tumor suppressors: getting a FERM grip on growth regulation.
        J Cell Sci. 2002; 115: 3991-4000
        • Yu H.
        • Zhang Y.
        • Ye L.
        • Jiang W.G.
        The FERM family proteins in cancer invasion and metastasis.
        Front Biosci. 2011; 16: 1536-1550
        • Wang J.
        • Cai Y.
        • Penland R.
        • Chauhan S.
        • Miesfeld R.L.
        • Ittmann M.
        Increased expression of the metastasis-associated gene Ehm2 in prostate cancer.
        Prostate. 2006; 66: 1641-1652
        • Yu H.
        • Ye L.
        • Mansel R.E.
        • Zhang Y.
        • Jiang W.G.
        Clinical implications of the influence of Ehm2 on the aggressiveness of breast cancer cells through regulation of matrix metalloproteinase-9 expression.
        Mol Cancer Res. 2010; 8: 1501-1512
        • Conway K.
        • Ruge F.
        • Price P.
        • Harding K.G.
        • Jiang W.G.
        Hepatocyte growth factor regulation: an integral part of why wounds become chronic.
        Wound Repair Regen. 2007; 15: 683-692
        • Zuker M.
        Mfold web server for nucleic acid folding and hybridization prediction.
        Nucleic Acids Res. 2003; 31: 3406-3415
        • Jiang W.G.
        • Grimshaw D.
        • Lane J.
        • Martin T.A.
        • Abounder R.
        • Laterra J.
        • et al.
        A hammerhead ribozyme suppresses expression of hepatocyte growth factor/scatter factor receptor c-MET and reduces migration and invasiveness of breast cancer cells (vol. 7, p. 2555, 2001).
        Clin Cancer Res. 2002; 8: 300-310
        • Jiang W.G.
        • Hiscox S.
        • Hallett M.B.
        • Scott C.
        • Horrobin D.F.
        • Puntis M.C.A.
        Inhibition of hepatocyte growth factor-induced motility and in-vitro invasion of human colon-cancer cells by gamma-linolenic acid.
        Br J Cancer. 1995; 71: 744-752
        • Parr C.
        • Watkins G.
        • Mansel R.E.
        • Jiang W.G.
        The hepatocyte growth factor regulatory factors in human breast cancer.
        Clin Cancer Res. 2004; 10: 202-211
        • Wegener J.
        • Keese C.R.
        • Giaever I.
        Electric cell–substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces.
        Exp Cell Res. 2000; 259: 158-166
        • Boukamp P.
        • Petrussevska R.T.
        • Breitkreutz D.
        • Hornung J.
        • Markham A.
        • Fusenig N.E.
        Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.
        J Cell Biol. 1988; 106: 761-771
        • Weaver A.M.
        • Heuser J.E.
        • Karginov A.V.
        • Lee W.L.
        • Parsons J.T.
        • Cooper J.A.
        Interaction of cortactin and N-WASp with Arp2/3 complex.
        Curr Biol. 2002; 12: 1270-1278
        • Drew P.
        • Posnett J.
        • Rusling L.
        The cost of wound care for a local population in England.
        Int Wound J. 2007; 4: 149-155
        • Schafer M.
        • Werner S.
        Cancer as an overhealing wound: an old hypothesis revisited.
        Nat Rev Mol Cell Biol. 2008; 9: 628-638
        • Chen L.
        • Hughes R.A.
        • Baines A.J.
        • Conboy J.
        • Mohandas N.
        • An X.
        Protein 4.1R regulates cell adhesion, spreading, migration and motility of mouse keratinocytes by modulating surface expression of beta1 integrin.
        J Cell Sci. 2011; 124: 2478-2487
        • Jensen P.V.
        • Larsson L.I.
        Actin microdomains on endothelial cells: association with CD44, ERM proteins, and signaling molecules during quiescence and wound healing.
        Histochem Cell Biol. 2004; 121: 361-369
        • Haas M.A.
        • Vickers J.C.
        • Dickson T.C.
        Rho kinase activates ezrin-radixin-moesin (ERM) proteins and mediates their function in cortical neuron growth, morphology and motility in vitro.
        J Neurosci Res. 2007; 85: 34-46
        • Okayama T.
        • Kikuchi S.
        • Ochiai T.
        • Ikoma H.
        • Kubota T.
        • Ichikawa D.
        • et al.
        Attenuated response to liver injury in moesin-deficient mice: impaired stellate cell migration and decreased fibrosis.
        Biochim Biophys Acta. 2008; 1782: 542-548
        • Chishti A.H.
        • Kim A.C.
        • Marfatia S.M.
        • Lutchman M.
        • Hanspal M.
        • Jindal H.
        • et al.
        The FERM domain: a unique module involved in the linkage of cytoplasmic proteins to the membrane.
        Trends Biochem Sci. 1998; 23: 281-282
        • Miki H.
        • Miura K.
        • Takenawa T.
        N-WASP, a novel actin-depolymerizing protein, regulates the cortical cytoskeletal rearrangement in a PIP2-dependent manner downstream of tyrosine kinases.
        EMBO J. 1996; 15: 5326-5335
        • Manchanda N.
        • Lyubimova A.
        • Ho H.Y.
        • James M.F.
        • Gusella J.F.
        • Ramesh N.
        • et al.
        The NF2 tumor suppressor Merlin and the ERM proteins interact with N-WASP and regulate its actin polymerization function.
        J Biol Chem. 2005; 280: 12517-12522
        • Suetsugu S.
        • Hattori M.
        • Miki H.
        • Tezuka T.
        • Yamamoto T.
        • Mikoshiba K.
        • et al.
        Sustained activation of N-WASP through phosphorylation is essential for neurite extension.
        Dev Cell. 2002; 3: 645-658