title: [RNA interference-mediated silencing of cytochrome P450 3A4 gene in CHL-3A4 cells].
abstract: OBJECTIVE
  To investigate the effect of small hairpin interfering RNA (shRNA) in suppressing cytochrome P450 3A4 (CYP3A4) gene expression in CHL-3A4 cells.
  
  
  METHODS
  Three shRNA expression vectors targeting CYP3A4 gene (CYP3A4 I, C YP3A4 II, and CYP3A4 III, respectively) were designed, synthesized and transfected into CHL-3A4 cells via liposomes. The inhibitory effect of shRNA on CYP 3A4 gene expression was detected by Western blotting and RT-PCR, and the effect of shRNA transfection in suppressing cyclophosphamide-induced cytotoxicity was measured using MTT assay.
  
  
  RESULTS
  The vector carrying CYP3A4 III shRNA significantly reduced the expression of CYP3A4 gene at both the mRNA (75%) and protein levels (80%) in CHL3A4 cells. The cytotoxicity of cyclophosphamide was markedly inhibited by CYP3A4 III-mediated suppression of CYP3A4 gene expression by 75% in CHL-3A4 cells.
  
  
  CONCLUSIONS
  The vector-mediated RNA interference can suppress CYP3A4 gene expression in CHL-3A4 cells, and RNA interference technique provides a new means for studying cytochrome P450 gene function in mammalian cells.
authors.affiliation: Department of Pharmacology, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Gui-zhong
authors.initials: GZ
authors.lastname: Yang
authors.forename: Hua-rong
authors.initials: HR
authors.lastname: Yu
authors.forename: Ye
authors.initials: Y
authors.lastname: Yuan
authors.forename: Qi-xin
authors.initials: QX
authors.lastname: Zhou
authors.forename: Ying-ju
authors.initials: YJ
authors.lastname: Liu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D006224:Cricetinae; D051544:Cytochrome P-450 CYP3A / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000166:cytology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human