Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression  

作　　者：Siyun Xu Yongsheng Xiao Li Li Lushan Yu Huidi Jiang Aiming Yu Su Zeng 

机构地区：[1]Department of Pharmaceutical Analysis and Drug Metabolism,College of Pharmaceutical Sciences,Zhejiang University,Hangzhou 310058,China [2]Department of Biochemistry&Molecular Medicine,UC-Davis Medical Center,Sacramento,CA 95817,USA

出　　处：《Acta Pharmaceutica Sinica B》2014年第5期350-357,共8页药学学报（英文版）

基　　金：This project was supported by the grants from National Natural Science Foundation of China(81173120,81302834);Major Program of National Natural Science Foundation of China(2012ZX09506001-004).

摘　　要：RNA interference(RNAi)is useful for selective gene silencing.Cytochrome P4503A4(CYP3A4),which metabolizes approximately 50% of drugs in clinical use,plays an important role in drug metabolism.In this study,we aimed to develop a short hairpin RNA(shRNA)to modulate CYP3A4 expression.Three new shRNAs(S1,S2 and S3)were designed to target the coding sequence(CDS)of CYP3A4,cloned into a shRNA expression vector,and tested in different cells.The mixture of three shRNAs produced optimal reduction(55%)in CYP3A4 CDS-luciferase activity in both CHL and HEK293 cells.Endogenous CYP3A4 expression in HepG2 cells was decreased about 50%at both mRNA and protein level after transfection of the mixture of three shRNAs.In contrast,CYP3A5 gene expression was not altered by the shRNAs,supporting the selectivity of CYP3A4 shRNAs.In addition,HepG2 cells transfected with CYP3A4 shRNAs were less sensitive to Ginkgolic acids,whose toxic metabolites are produced by CYP3A4.These results demonstrate that vector-based shRNAs could modulate CYP3A4 expression in cells through their actions on CYP3A4 CDS,and CYP3A4 shRNAs may be utilized to define the role of CYP3A4 in drug metabolism and toxicity.

关 键 词：RNAi Cytochrome P450 CYP3A4 SHRNA CHEMOSENSITIVITY 

分 类 号：R96[医药卫生—药理学]