title: Gene expression analysis of an epidermolysis bullosa simplex Dowling-Meara cell line by subtractive hybridization: recapitulation of cellular differentiation, migration and wound healing.
abstract: An intact keratin 5/keratin 14 intermediate filament cytoskeleton is vital for the integrity of basal keratinocytes and for the development and maintenance of epidermal structures. In patients with epidermolysis bullosa simplex Dowling-Meara (EBS-DM), heterozygous mutations in the keratin 14 gene in keratinocytes cause a cytoskeletal collapse leading to fragile cells susceptible to cellular stress. The primary aim of this work was to extend analysis of differentially expressed genes in an EBS-DM model cell line to obtain insights into the molecular consequences resulting from the keratin 14 mutation. In a first step, suppression subtractive hybridization (SSH), a powerful technology to enrich for differentially expressed genes, was used to identify genes whose up-regulation may be a direct or indirect result of the keratin 14 mutation, R125P. We discovered 55 candidate genes (SSH genes) that were further analysed by RTq-PCR. Of the 55 SSH genes, 14 (25.45%) were found to be congruently up-regulated. Bioinformatic analysis revealed significant enrichment of genes regulating epidermal development, migration, apoptosis and wound healing.
authors.affiliation: Division of Molecular Dermatology, Department of Dermatology, Paracelsus Private Medical University Salzburg, Salzburg, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Wagner
authors.forename: Helmut
authors.initials: H
authors.lastname: Hintner
authors.forename: Johann W
authors.initials: JW
authors.lastname: Bauer
authors.forename: Kamil
authors.initials: K
authors.lastname: Onder
mesh_terms: D019943:Amino Acid Substitution / Q000502:physiology; D051017:Apoptosis Regulatory Proteins / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics*; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000235:genetics*; D019295:Computational Biology / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D016110:Epidermolysis Bullosa Simplex / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D053547:Keratin-14 / Q000235:genetics; D015603:Keratinocytes / Q000378:metabolism*; D007633:Keratins / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics; D014945:Wound Healing / Q000235:genetics*
chemical_list: D051017:Apoptosis Regulatory Proteins; D018076:DNA, Complementary; C507540:KRT14 protein, human; D053547:Keratin-14; D012269:Ribosomal Proteins; D007633:Keratins