Gene expression analysis of an epidermolysis bullosa simplex Dowling-Meara cell line by subtractive hybridization: recapitulation of cellular differentiation, migration and wound healing.

[epidermolysis bullosa simplex]

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 , R 125 P . 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 .