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.

Attached Files
Name Description MIMEType Size Downloads

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, 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
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
Persistent URL http://hdl.handle.net/10536/DRO/DU:30009156

Document type: Journal Article
Collection: School of Medicine
Connect to link resolver
 
Unless expressly stated otherwise, the copyright for items in DRO is owned by the author, with all rights reserved.

Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 8 times in TR Web of Science
Scopus Citation Count Cited 11 times in Scopus
Google Scholar Search Google Scholar
Access Statistics: 452 Abstract Views, 0 File Downloads  -  Detailed Statistics
Created: Mon, 13 Oct 2008, 15:52:47 EST

Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.