[1]卢忠燕,甘立霞,王哲,等.抑制FoxO1基因表达的siRNA重组腺病毒载体的构建及功能鉴定[J].陆军军医大学学报(原第三军医大学学报),2007,29(23):2215-2218.
　LU Zhong-yan,GAN Li-xia,WANG Zhe,et al.Recombinant adenovirus vector construction of siRNA targeting FoxO1 and its functional characterization[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(23):2215-2218.
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陆军军医大学学报（原第三军医大学学报）[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第23期
页码:
2215-2218
栏目:
论著
出版日期:
2007-12-15

文章信息/Info

Title:
Recombinant adenovirus vector construction of siRNA targeting FoxO1 and its functional characterization
作者:
卢忠燕; 甘立霞; 王哲; 何凤田; 邹全明; 曹廷兵
第三军医大学:基础医学部生物化学与分子生物学教研室，医学检验系临床微生物学及免疫学教研室，大坪医院野战外科研究所内分泌科，重庆市高血压病研究所
Author(s):
LU Zhong-yan;  GAN Li-xia;  WANG Zhe;  HE Feng-tian;  ZOU Quan-ming;  CAO Ting-bing
Department of Biochemistry and Molecular Biology, College of Medicine, Department of Clinical Microbiology, College of Medical Laboratory, Institute of Hypertension Research, Institute of Surgery Research, Daping Hospital, Third Military Medical University
关键词:
FoxO1;  siRNA;  腺病毒载体;  HepG2细胞
Keywords:
FoxO1;  siRNA;  adenoviral vector;  HepG2 cell line
分类号:
R394-33; R394.2; R735.7
文献标志码:
A
摘要:
目的  构建针对转录因子FoxO1的siRNA腺病毒载体，并鉴定重组腺病毒在人肝癌细胞系HepG2中对内源性FoxO1基因表达的影响。  方法  设计并合成针对FoxO1的siRNA的靶DNA序列，克隆于穿梭载体pAdTrack-CMV中，与腺病毒骨架质粒pAdeasy-1在BJ5183细菌中进行同源重组，转染293细胞，包装得到含siFoxO1的重组腺病毒，体外转染人肝癌细胞系HepG2，Western blot检测FoxO1蛋白表达水平的变化。  结果  成功构建针对FoxO1的siRNA重组腺病毒载体，该重组腺病毒能显著抑制HepG2细胞中FoxO1蛋白的表达(P<0.01)。  结论  成功构建了针对FoxO1的siRNA重组腺病毒载体，能有效抑制HepG2细胞中FoxO1的表达。
Abstract:
Objective  To construct an adenovirus vector that expresses small interfering RNA (siRNA) against FoxO1 to shutdown its gene expression.   Methods  The FoxO1 template DNA sequence was designed by using online tools. Then the sense and antisense siRNA oligonucleotide templates were chemically synthesized, annealed and cloned into adenoviral shuttle vector pAdTrack-CMV. The recombinant adenovirus vector pAd-si-FoxO1 was obtained by homologous recombination with pAdTrack-CMV and pAdeasy-1 in bacteria BJ5183. The recombined adenovirus was produced in 293 cells and subsequently infected HepG2 cells. The FoxO1 protein levels were detected by Western blot.   Results  The adenovirus vector expressing small interfering RNA for FoxO1 gene could specifically shutdown the endogenous FoxO1 protein in HepG2 cells.    Conclusion  The pAd-si-FoxO1 can effectively downregulate FoxO1 expression in HepG2 cell line.

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更新日期/Last Update: 2008-05-20