| RRC ID |
38280
|
| 著者 |
Sato W, Suzuki H, Sasaki T, Kumagai T, Sakaguchi S, Mizugaki M, Miyairi S, Yamazoe Y, Nagata K.
|
| タイトル |
Construction of a system that simultaneously evaluates CYP1A1 and CYP1A2 induction in a stable human-derived cell line using a dual reporter plasmid.
|
| ジャーナル |
Drug Metab Pharmacokinet
|
| Abstract |
Human CYP1A1 and CYP1A2 genes are in a head-to-head orientation on chromosome 15 and are separated by a 23-kb intergenic space. To our knowledge, this is the first report on a stable cell line that contains the 23-kb full-length regulatory region and is able to simultaneously assess the transcriptional activation of CYP1A1 and CYP1A2 genes. The stable cell line that constitutively expresses the reporter activities was constructed by inserting the dual reporter plasmid containing the 23-kb region between the CYP1A1 and CYP1A2 genes into the chromosome. Transcriptional activation of the CYP1A1 and CYP1A2 genes was measured simultaneously using luciferase (Luc) and secreted alkaline phosphatase (SEAP) activities, respectively. To demonstrate the utility of the stable cell line, CYP1A1/1A2 induction by the majority of compounds previously identified as CYP1A1/1A2 inducers was measured. The results clearly show that all compounds caused induction of reporter activities. In addition to assessing transcriptional activation of the CYP1A1 and CYP1A2 genes by measuring reporter activities, we determined the intrinsic CYP1A1 and CYP1A2 mRNA levels by treating them with the same compounds. The results suggest that this stable cell line may be used to rapidly and accurately predict CYP1A1/1A2 induction.
|
| 巻・号 |
25(2)
|
| ページ |
180-9
|
| 公開日 |
2010-1-1
|
| DOI |
10.2133/dmpk.25.180
|
| PII |
JST.JSTAGE/dmpk/25.180
|
| PMID |
20460824
|
| MeSH |
Cell Line
Chromosomes, Human, Pair 15
Cytochrome P-450 CYP1A1 / biosynthesis
Cytochrome P-450 CYP1A1 / genetics*
Cytochrome P-450 CYP1A1 / metabolism
Cytochrome P-450 CYP1A2 / biosynthesis
Cytochrome P-450 CYP1A2 / genetics*
Cytochrome P-450 CYP1A2 / metabolism
Enzyme Induction
Gene Expression Regulation, Enzymologic
Genes, Reporter / genetics*
Genetic Engineering / methods*
Genetic Vectors
Humans
Luciferases
Multigene Family
Plasmids / genetics*
RNA, Messenger / metabolism*
Recombination, Genetic
Transcriptional Activation / genetics
Transfection
|
| IF |
2.772
|
| 引用数 |
10
|
|
WOS 分野
|
PHARMACOLOGY & PHARMACY
|
| リソース情報 |
| ヒト・動物細胞 |
Hep G2(RCB1648) |
