Development of a whole-cell screening system for evaluation of the human CYP1A2-mediated metabolism
| Autor: | Hsien-Tsung Yao, Kao-Lu Pan, John T.A. Hsu, Chih-Chieh Chu |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Berberine
Drug-Related Side Effects and Adverse Reactions Cytochrome P-450 CYP1A2 Inhibitors Cell Drug Evaluation Preclinical Bioengineering Applied Microbiology and Biotechnology Cell Line Green fluorescent protein Theophylline Cytochrome P-450 CYP1A2 medicine Humans Enzyme Inhibitors Cytotoxicity biology CYP1A2 Cytochrome P450 Recombinant Proteins Internal ribosome entry site medicine.anatomical_structure Biochemistry Cell culture biology.protein Drug metabolism Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 108:2932-2940 |
| ISSN: | 0006-3592 |
| DOI: | 10.1002/bit.23256 |
| Popis: | Cytochrome P450 1A2 (CYP1A2) is an important member of cytochrome P450 involved in drug metabolism. In this study, a cell line, Huh7-1A2-I-E, with high expression level of CYP1A2 is established based on Huh7 cells. To achieve this, we constructed a recombinant lentiviral vector, pLenti-1A2-I-E, containing a single promoter encoding CYP1A2 followed by an internal ribosome entry site (IRES) to permit the translation of enhanced green fluorescence protein (EGFP). Such a design has greatly facilitated the selection of stable cell lines because the translations of CYP1A2 and EGFP proteins would be based on a single bi-cistronic mRNA. The Huh7-1A2-I-E cells were evaluated as a cell-based model for identification of CYP1A2 inhibitors and for studies of cytotoxicity resulted from CYP-mediated drug metabolism. Treatment of Huh7-1A2-I-E cells and the Huh7-E control cells with aflatoxin B1 showed that cells with CYP1A2 expression are much more sensitive to aflatoxin B1 and the cellular toxicity of aflatoxin B1 in Huh7-1A2-I-E cells could be prevented by furafylline, a CYP1A2 inhibitor. A collection of approximately 200 drugs were screened using this system and results indicate that for most drugs the metabolism by CYP1A2 is unlikely to have made a major contribution to the in vitro cytotoxicity except for thimerosal and evoxine. Several previously unidentified CYP1A2 inhibitors such as evoxine and berberine were also identified in this study. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text