[RNA interference-mediated silencing of cytochrome P450 3A4 gene in CHL-3A4 cells].
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.
Yang
Gui-zhong
GZ
Department of Pharmacology, Chongqing Medical University, Chongqing 400016, China.
Yu
Hua-rong
HR
Yuan
Ye
Y
Zhou
Qi-xin
QX
Liu
Ying-ju
YJ
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*
D034741:RNA, Small Interfering; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human