Gene expression analysis of EpiDerm following exposure to SLS using cDNA microarrays.
There is a need to investigate the mechanistic basis of the human skin irritation response if relevant in vitro test systems for the predictive identification of skin irritation hazards are to be developed. Recent progress in genomics technologies mean that tools for the identification and investigation of important biochemical events in the processes of skin irritation are now available. The aim of this work was to identify genes (for further mechanistic investigation) which may be regulated in response to skin irritation, following exposure of the EpiDerm skin model to the known skin irritant sodium lauryl sulphate (SLS). EpiDerm cultures were treated in triplicate with a non-cytotoxic dose of SLS (0.1 mg/ml, as determined by the MTT assay and histological examination) for 15 min, 30 min, 1 h, 2 h, 3 h, 4 h and 24 h. Total RNA was extracted from the pooled EpiDerm cultures and used to probe Atlas human arrays (Clontech) covering approximately 3600 genes. Preliminary data indicated an up-regulation at early time points (15-30 min) of a number of genes involved in transportation (e.g. the sodium and chloride dependent taurine transporter) and receptors (e.g. ZAP70 and protocadherin 42 precursor). The gene encoding the UV excision repair protein and other DNA repair genes (e.g. DNA-directed RNA polymerase II) were up-regulated after 1-3 h, along with TGF beta 3 and other tumour suppressors, which play a role in cellular development and wound healing. At the later time points of 4-24 h, genes involved in protein translation (e.g. Cathepsin D receptor) and metabolism (e.g. CYP27A) were up-regulated. In addition, a number of genes were down-regulated in response to treatment with SLS, although these followed less of a time dependent pattern. These results indicate the differential regulation of a number of genes in response to treatment with SLS, some of which may provide additional clues to the molecular events underpinning the irritation response to this particular surfactant and possibly to other chemical irritants.
Fletcher
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ST
SEAC Toxicology Unit, Unilever Research, Sharnbrook, Bedfordshire MK44 1LQ, UK. samantha.fletcher@unilever.com
Baker
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VA
Fentem
J H
JH
Basketter
D A
DA
Kelsell
D P
DP
D000826:Animal Testing Alternatives; D002478:Cells, Cultured; D017453:Dermatitis, Irritant / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D020869:Gene Expression Profiling; D023281:Genomics; D006801:Humans; D007509:Irritants / Q000633:toxicity*; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000032:analysis; D012867:Skin / Q000187:drug effects*; D012967:Sodium Dodecyl Sulfate / Q000633:toxicity*; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism; D013997:Time Factors; D015854:Up-Regulation
D007509:Irritants; D013778:Tetrazolium Salts; D013844:Thiazoles; D012967:Sodium Dodecyl Sulfate; D012313:RNA; C022616:thiazolyl blue