title: Regulation of cytochrome P450 2E1 under hypertonic environment through TonEBP in human hepatocytes.
abstract: Whereas the liver as well as the other organs are continually exposed to the change of osmotic status, it has never been investigated whether activities and gene expressions of drug-metabolizing enzymes, including cytochromes P450, are dependent on osmotic change in the liver. In the present study, we determined that CYP2E1 is induced under hypertonic environments at a transcriptional level in human primary hepatocytes, as assessed by cDNA microarray and real time-reverse transcription-polymerase chain reaction analyses. Both a protein level and the catalytic activity of CYP2E1 were consistently increased in response to hypertonic conditions. In promoter-reporter assay, it was demonstrated that -586 to -566 in the CYP2E1 5'-flanking region was necessary for 2E1 promoter activation by hypertonic stimulation. It is noteworthy that tonicity-response element (TonE) consensus sequence was found at -578 to -568 in human CYP2E1 5'-flanking region, and electrophoretic mobility shift assay demonstrated the interaction of TonE binding protein (TonEBP) with TonE motif of CYP2E1 promoter. Furthermore, cotransfection of a CYP2E1 promoter construct with wild-type TonEBP expression vector enhanced promoter activity under both isotonic and hypertonic conditions, whereas dominant-negative TonEBP suppressed an induction of CYP2E1 promoter activity. These results indicate that the level of CYP2E1 is induced by hypertonic condition via TonEBP transactivation. The present study suggests that osmotic status may influence individual responses to the substrate of CYP2E1.
authors: Department of Clinical Pharmacology and Pharmacogenomics, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors: Takashi
authors: T
authors: Ito
authors: Koko
authors: K
authors: Asakura
authors: Katsuhiko
authors: K
authors: Tougou
authors: Tsuyoshi
authors: T
authors: Fukuda
authors: Ryuji
authors: R
authors: Kubota
authors: Shinpei
authors: S
authors: Nonen
authors: Yasushi
authors: Y
authors: Fujio
authors: Junichi
authors: J
authors: Azuma
mesh_terms: D002478:Cells, Cultured; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics; D019392:Cytochrome P-450 CYP2E1 / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D022781:Hepatocytes / Q000201:enzymology*; D006801:Humans; D006982:Hypertonic Solutions; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000502:physiology*
chemical_list: D006982:Hypertonic Solutions; C118125:NFAT5 protein, human; D014157:Transcription Factors; D019392:Cytochrome P-450 CYP2E1; D019389:Cytochrome P-450 CYP2D6