Small interfering RNA mediated silencing of cytochrome P450 3A4 gene
- VernacularTitle:RNA干扰抑制CYP3A4基因表达的实验研究
- Author:
Jie CHEN
;
Lizi ZHAO
;
Ying DENG
;
Xueding WANG
;
Su GUAN
;
Min HUANG
- Publication Type:Journal Article
- Keywords:
RNA interference;
Cytochrome P450;
Gene expression;
Cyclophosphamide;
Laboratory techniques and procedures
- From:
Chinese Journal of Primary Medicine and Pharmacy
2006;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibitory effect of the cytochrome P450 3A4(CYP3A4) gene expression and function in CHL-3A4 cells lines by vector-ecpressed small hairpin interfering RNA(shRNA).Methods The shRNA expression vectors targeting CYP 3A4 gene(CYP3A4Ⅰ,CYP3A4Ⅱ,CYP3A4Ⅲ) were designed and constructed.The inhibitory effect of shRNA was detected by Western blot and RT-PCR analysis.The inhibitory effect of cytotoxic of cyclophosphamide using shRNA was measured by MTT assay.Results CYP3A4Ⅲ shRNA expressing vector significantly reduced the CYP3A4 mRNA(70%) and protein expression levels(75%) of the CYP3A4 gene in CHL3A4 cells suppression of CYP3A4 gene expression by CYP3A4Ⅲ largely reversed cyclophosphamide cytotoxicity(75%) in CHL-3A4 cells.Conclusion Vector-based RNAi could suppress CYP3A4 gene expression and function,and the use of RNAi to inhibit CYP3A4 gene expression in mammalian cells was a promising new tool for the study of cytochrome P450 gene function.
