Small interfering RNA-mediated silencing of cytochrome P450 3A4 gene

Jie Chen, Xiao Xia Yang, Min Huang, Ze Ping Hu, Ming He, Wei Duan, Eli Chan, Fwu Shan Sheu, Xiao Chen, Shu Feng Zhou

Research output: Contribution to journalArticle

11 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1650-1657
Number of pages8
JournalDrug Metabolism and Disposition
Volume34
Issue number9
DOIs
StatePublished - Aug 31 2006

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

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  • Cite this

    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. https://doi.org/10.1124/dmd.106.009837