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KMID : 0604220070140010017
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Korean Journal Investigative Dermatology
2007 Volume.14 No. 1 p.17 ~ p.28
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Identification of the Genes Differentially Expressed in the Keratinocytes by Treatment with 12-O-Tetradecanoylphorbol-13-Acetate Using Suppression Subtractive Hybridization
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Lee Joo-Hee
Lee Jeung-Hoon
Jeon Eun-Kyung
Song Hae-Jun
Seo Eun-Young
Kim Chang-Duk
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Abstract
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TPA (12-O-Tetradecanoylphorbol-13-Acetate), a potent tumor promoter, is well known to accelerate the differentiation of cultured human keratinocytes. So far many studies have been done to uncover the genes expressed in keratinocytes by TPA treatment. But by technical reason, there were limitations to identify specific genes differentially expressed by TPA treatment on a large scale. Suppression subtractive hybridization (SSH) enabling efficient amplification of the specific gene differentially expressed in the cell was used. Cultured human foreskin keratinocytes were treated with TPA and 795 differentially expressed clones were randomly retrieved. By reverse northern blot analyses 237 clones were found to be over-expressed. After sequencing, they were shown to represent 29 different genes, of which all but one were confirmed as differentially expressed by Northern blot analysis. Of those, 10 genes are already known to have a role in keratinocytes. Nineteen genes, and 7 EST genes were not known in keratinocytes. These differentially expressed genes may provide significant opportunities for further understanding epidermal keratinocyte differentiation and carcinogenesis.
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KEYWORD
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Keratinocyte differentiation, TPA, Suppression subtractive hybridization (SSH)
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