Characterization of a Human Keratinocyte HaCaT Cell Line Model to Study the Regulation of CYP2S1

• DOI: 10.1124/dmd.111.042085
• Publication Date: 2011-10-29T02:42:47Z
• Authors: Alison D. McNeilly; Julie A. Woods; Sally H. Ibbotson; C. Roland Wolf; Gillian Smith
• Keywords: HaCaT

CYP2S1 is an extrahepatic cytochrome P450 (P450) that shows marked individuality in constitutive and inducible expression. mRNA expression increased psoriasis by treatments for psoriasis, including retinoids UV radiation, although endogenous substrates remain poorly characterized. Because previous model systems have overexpressed modified bacteria, human HaCaT keratinocyte cells were screened regulatable activity compared with a novel Chinese hamster ovary (CHO)-based cell line engineered to stably coexpress NADPH reductase. Constitutive additional P450s, retinoid acid receptors (RARα, RARβ, RARγ), X (RXRα, RXRβ RXRγ) was assessed quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis cells. Cells then exposed or radiation (UVR), changes abundance further examined qRT-PCR analysis. similar skin, abundant RARα RARγ (but not RARβ) all RXR isoforms also detectable. All-<i>trans</i> retinoic (atRA) induced CYPS1 more potently than 9-<i>cis</i> RA 13-<i>cis</i> RA. P450-dependent atRA metabolism demonstrated cells, very metabolite profile produced our CYP2S1-expressing CHO UVR, CYP1B1, known UVR-inducible P450. Our results demonstrate functional thus identifying utility of regulation substrate specificity.