Evaluation of the skin irritation using a DNA microarray on a reconstructed human epidermal model.
To avoid the need to use animals to test the skin irritancy potential of chemicals and cosmetics, it is important to establish an in vitro method based on the reconstructed human epidermal model. To evaluate skin irritancy efficiently and sensitively, we determined the gene expression induced by a topically-applied mild irritant sodium dodecyl sulfate (SDS) in a reconstructed human epidermal model LabCyte EPI-MODEL (LabCyte) using a DNA microarray carrying genes that were related to inflammation, immunity, stress and housekeeping. The expression and secretion of IL-1alpha in reconstructed human epidermal culture is known to be induced by irritation. We detected the induction of IL-1alpha expression and its secretion into the cell culture medium by treatment with 0.075% SDS for 18 h in LabCyte culture using DNA microarray, quantitative reverse-transcription polymerase chain reaction (RT-PCR) and ELISA. DNA microarray analysis indicated that the expression of 10 of the 205 genes carried on the DNA microarray was significantly induced in a LabCyte culture by 0.05% or 0.075% SDS irritation for 18 h. RT-PCR analysis confirmed that SDS treatment significantly induced the expressions of interleukin-1 receptor antagonist (IL-1RN), FOS-like antigen 1 (FOSL1), heat shock 70 kDa protein 1A (HSPA1) and myeloid differentiation primary response gene (88) (MYD88), as well as the known marker genes for irritation IL-1beta and IL-8 in a LabCyte culture. Our results showed that a DNA microarray is a useful tool for efficiently evaluating mild skin irritation using a reconstructed human epidermal model.
Niwa
Makoto
M
Katakura Chikkarin Co., Ltd, Ibaraki, Japan.
Nagai
Kanji
K
Oike
Hideaki
H
Kobori
Masuko
M
D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D000078404:Epidermal Cells; D004817:Epidermis / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007509:Irritants / Q000633:toxicity*; D053594:Myeloid Differentiation Factor 88 / Q000096:biosynthesis / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D016760:Proto-Oncogene Proteins c-fos / Q000096:biosynthesis / Q000235:genetics; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D017472:Receptors, Interleukin-1 / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000187:drug effects* / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000633:toxicity; D013778:Tetrazolium Salts; D013844:Thiazoles; D046509:Tissue Culture Techniques; D000099169:Fos-Related Antigen 1
D004247:DNA; D018840:HSP70 Heat-Shock Proteins; D007509:Irritants; D053594:Myeloid Differentiation Factor 88; D016760:Proto-Oncogene Proteins c-fos; D012313:RNA; D017472:Receptors, Interleukin-1; D012967:Sodium Dodecyl Sulfate; D013778:Tetrazolium Salts; D013844:Thiazoles; D000099169:Fos-Related Antigen 1; C022616:thiazolyl blue