Background:Keloids are an extreme form of abnormal scarring that result from a pathological fibroproliferative wound healing process.The molecular mechanisms driving keloid pathology remain incompletely understood,hindering development of targeted,effective therapies.Recent studies in our laboratory demonstrated that keloid keratinocytes exhibit adhesion abnormalities and display a transcriptional signature reminiscent of cells undergoing epithelial-mesenchymal transition(EMT),suggesting a role for EMT in keloid pathology.In the current study,we further define the EMT-like phenotype of keloid scars and investigate regulation of EMT-related genes in keloid.Methods:Primary keratinocytes from keloid scar and normal skin were cultured in the presence or absence of transforming growth factor beta 1(TGF-β1)+/-inhibitors of TGF-β1 and downstream signaling pathways.Gene expression was measured using quantitative polymerase chain reaction.Migration was analyzed using an in vitro wound healing assay.Proteins in keloid scar and normal skin sections were localized by immunohistochemistry.Statistical analyses utilized SigmaPlot(SyStat Software,San Jose,CA)or SAS?(SAS Institute,Cary,NC).Results:In keloid and normal keratinocytes,TGF-β1 regulated expression of EMT-related genes,including hyaluronan synthase 2,vimentin,cadherin-11,wingless-type MMTV integration site family,member 5A,frizzled 7,ADAM metallopeptidase domain 19,and interleukin-6.Inhibition of canonical TGF-β1 signaling in keloid keratinocytes significantly inhibited expression of these genes,and TGF-β1 stimulation of normal keratinocytes increased their expression.The inhibition of the extracellular signal-regulated kinase 1/2(ERK1/2)signaling pathway or the p38 mitogen-activated protein kinase pathway attenuated TGF-β1-induced expression of subsets of these genes.Migration of keloid keratinocytes,previously shown to be increased compared with normal keratinocytes,was significantly reduced by inhibition of TGF-β1 or ERK1/2 signaling.Biomarkers of EMT,includi
Jennifer M.HahnKevin L.McFarlandKelly A.CombsDorothy M.Supp
