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Title: Small interfering RNA-mediated silencing of cytochrome P450 3A4 gene
Authors: Chen, J.
Yang, X.-X.
Huang, M.
Hu, Z.-P.
He, M.
Duan, W.
Chan, E. 
Sheu, F.-S. 
Chen, X.
Zhou, S.-F. 
Issue Date: 2006
Citation: Chen, J., Yang, X.-X., Huang, M., Hu, Z.-P., He, M., Duan, W., Chan, E., Sheu, F.-S., Chen, X., Zhou, S.-F. (2006). Small interfering RNA-mediated silencing of cytochrome P450 3A4 gene. Drug Metabolism and Disposition 34 (9) : 1650-1657. ScholarBank@NUS Repository.
Abstract: RNA interference (RNAi) is a specific and powerful tool used to manipulate gene expression and study gene function. The cytochrome P450 3A4 (CYP3A4) can metabolize more than 50% of drugs. In the present study, we investigated whether vector-expressed small interfering RNAs (siRNAs) altered the CYP3A4 expression and function using the Chinese hamster cell line (V79) overexpressing CYP3A4 (CHL-3A4). Three different siRNA oligonucleotides (3A4I, 3A4II, and 3A4III) were designed and tested for their ability to interfere with CYP3A4 gene expression. Our study demonstrated that transient transfection of CHL-3A4 cells with the 3A4III siRNAs, but not 3A4I and II, significantly reduced CYP3A4 mRNA levels by 65% and protein expression levels by 75%. All these siRNAs did not affect the expression of CYP3A5 at both mRNA and protein levels in V79 cells overexpressing CYP3A5. Transfection of CHL-3A4 cells with 3A4III siRNAs significantly diminished the cytotoxicity of two CYP3A4 substrate drugs, cyclophosphamide and ifosfamide, in CHL-3A4 cells, with the IC50 increased from 55 to 210 μM to >1000 μM. Nifedipine at 5.78, 14.44, and 28.88 μM was significantly (P < 0.01) depleted by approximately 100, 40, and 22%, respectively, in S9 fractions from CHL-3A4 cells compared with parental CHL-pIC19h cells. In addition, transfection of the CHL-3A4 cells with vectors expressing the 3A4III siRNAs almost completely inhibited CYP3A4-mediated nifedipine metabolism. This study demonstrated, for the first time, the specific suppression of CYP3A4 expression and function using vector-based RNAi technique. The use of RNAi is a promising tool for the study of cytochrome P450 family function. Copyright © 2006 by The American Society for Pharmacology and Experimental Therapeutics.
Source Title: Drug Metabolism and Disposition
ISSN: 00909556
DOI: 10.1124/dmd.106.009837
Appears in Collections:Staff Publications

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