| RRC ID |
71911
|
| 著者 |
Shindo S, Shioya A, Watanabe M, Sasaki T, Suzuki H, Kumagai T, Hwang GW, Nagata K.
|
| タイトル |
Development of an adenovirus-mediated reporter assay system to detect a low concentration of retinoic acid in MCF-7 cells.
|
| ジャーナル |
J Toxicol Sci
|
| Abstract |
Retinoic acid, an active form of vitamin A, plays very important roles in mammalian embryogenesis. The concentration of retinoic acid is extremely low and strictly regulated by enzymes of cytochrome P450 (CYP) family, CYP26s (CYP26A1, CYP26B1 and CYP26C1) in the cells. Therefore, it is thought that changes in CYP26s activities due to exposure to a wide variety of drugs and chemicals exhibit teratogenicity. In this study, to easily detect the changes in retinoic acid level, we constructed an adenovirus-mediated reporter assay system using the promoter region of the CYP26A1 gene and inserting retinoic acid response element (RARE) and retinoid X response element (RXRE) into the downstream of the luciferase gene of reporter plasmid, which highly increased the response to retinoic acid. Reporter activity significantly increased in a concentration-dependent manner with retinoic acid; this increase was also observed at least after treatment with a very low concentration of 1 nM retinoic acid. This increase was suppressed by the accelerated metabolism of retinoic acid due to the overexpression of CYP26A1; however, this suppression was almost completely suspended by treatment with talarozole, a CYP26 inhibitor. In conclusion, the reporter assay system constructed using the induction of CYP26A1 expression is a risk assessment system that responds to extremely low concentrations of retinoic acid and is useful for assessing the excess vitamin A mediated teratogenicity caused by various chemicals at the cellular level.
|
| 巻・号 |
47(6)
|
| ページ |
249-255
|
| 公開日 |
2022-1-1
|
| DOI |
10.2131/jts.47.249
|
| PMID |
35650141
|
| MeSH |
Adenoviridae* / genetics
Cytochrome P-450 Enzyme System / metabolism
Genes, Reporter
Humans
MCF-7 Cells
Promoter Regions, Genetic
Retinoic Acid 4-Hydroxylase / genetics
Teratogens* / analysis
Tretinoin* / analysis
Vitamin A
|
| IF |
1.737
|
| リソース情報 |
| ヒト・動物細胞 |
MCF7(RCB1904)
HeLa(RCB0007)
Hep G2(RCB1648)
Hep G2(RCB1886) |
