Adenovirus Vector-Mediated Reporter System forIn Vivo Analyses of Human CYP3A4 GeneActivation
| Autor: | Miki Shimada, Toru Yamazaki, Kiyoshi Nagata, Takayuki Okubo, Yasushi Yamazoe, Masayuki Furukawa, Makoto Ogino |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Male
Transcriptional Activation Genetic Vectors Gene Expression Biology Transfection medicine.disease_cause Biochemistry Dexamethasone Adenoviridae Mixed Function Oxygenases Viral vector Mice Cytochrome P-450 Enzyme System Genes Reporter In vivo Tumor Cells Cultured medicine Animals Cytochrome P-450 CYP3A Humans Clotrimazole Enzyme Inhibitors Luciferases Promoter Regions Genetic Glucocorticoids Molecular Biology Gene Regulation of gene expression Reporter gene Mutation Promoter General Medicine Molecular biology Growth Inhibitors Gene Expression Regulation Liver Bioreporter Rifampin |
| Zdroj: | Journal of Biochemistry. 131:71-78 |
| ISSN: | 0021-924X |
| DOI: | 10.1093/oxfordjournals.jbchem.a003079 |
| Popis: | The use of cultured mammalian cells and artificial promoters for analyses of gene regulation gives results that are sometimes inconsistent with in vivo events and thus inconclusive. To understand the in vivo mechanism of chemically mediated CYP3A4 gene activation, we have used a natural promoter of the CYP3A4 gene and an adenovirus as a reporter vector. The adenovirus reporter vector (AdCYP3A4-362) was constructed with a proximal promoter region (-362 to +11 nt) of the CYP3A4 gene and a luciferase-reporter gene. AdCYP3A4-362 was then infected into mice, and both the reporter and mouse CYP3A activities were measured. Clear increases in the reporter activity were observed in livers of all mice treated with chemicals. The profile of the CYP3A4 gene activation with chemicals was in good agreement with that of endogenous mouse CYP3A-mediated testosterone 6beta-hydroxylase. Introduction of nucleotide mutations in the receptor-binding region (ER-6) of the CYP3A4 promoter resulted in diminished reporter activity. These results indicate the advantage of the adenovirus-mediated in vivo system over the currently available in vitro systems for gene transcriptional activation. |
| Databáze: | OpenAIRE |
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