Small interfering RNA-mediated silencing of cytochromeP450 3A4 gene

Chen, Jie, Yang, Xiao-Xia, Huang, Min, Hu, Ze-Ping, He, Ming, Duan, Wei, Chan, Eli, Sheu, Fwu-Shan, Chen, Xiao and Zhou, Shu-Feng 2006, Small interfering RNA-mediated silencing of cytochromeP450 3A4 gene, Drug metabolism and disposition, vol. 34, no. 9, pp. 1650-1657, doi: 10.1124/dmd.106.009837.

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Title Small interfering RNA-mediated silencing of cytochromeP450 3A4 gene
Author(s) Chen, Jie
Yang, Xiao-Xia
Huang, Min
Hu, Ze-Ping
He, Ming
Duan, WeiORCID iD for Duan, Wei
Chan, Eli
Sheu, Fwu-Shan
Chen, Xiao
Zhou, Shu-Feng
Journal name Drug metabolism and disposition
Volume number 34
Issue number 9
Start page 1650
End page 1657
Publisher American Society for Pharmacology and Expreimental Therapeutics
Place of publication Bethesda, Md.
Publication date 2006-06-07
ISSN 0090-9556
Summary 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.
Language eng
DOI 10.1124/dmd.106.009837
Field of Research 111599 Pharmacology and Pharmaceutical Sciences not elsewhere classified
HERDC Research category C1.1 Refereed article in a scholarly journal
Copyright notice ©2006, The American Society for Pharmacology and Experimental Therapeutics
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Document type: Journal Article
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