Suppression of tissue inhibitor of metalloproteinase-1 by recombinant adeno-associated viruses carrying siRNAs in hepatic stellate cells

被引：15

作者：

Cong, Min ^{[1
]}

Liu, Tianhui ^{[1
]}

Wang, Ping ^{[1
]}

Xu, Yong ^{[1
]}

Tang, Shuzhen ^{[1
]}

Wang, Baoen ^{[1
]}

Jia, Jidong ^{[1
]}

Liu, Yong ^{[2
]}

Hermonat, Paul L. ^{[2
]}

You, Hong ^{[1
]}

机构：

[1] Capital Med Univ, Liver Res Ctr, Beijing Friendship Hosp, Beijing 100050, Peoples R China

[2] Univ Arkansas Med Sci, Dept Internal Med, Gene Therapy Ctr, Little Rock, AR 72205 USA

来源：

INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE | 2009年 / 24卷 / 05期

基金：

国家高技术研究发展计划(863计划);

关键词：

adeno-associated virus; small interfering RNA; tissue inhibitor of metalloproteinase-1; GENE-TRANSFER; VIRAL VECTORS; BEHAVIORAL RECOVERY; LIVER FIBROSIS; HEMOPHILIA-B; MOUSE MODEL; FACTOR-IX; IN-VIVO; DELIVERY; THERAPY;

D O I：

10.3892/ijmm_00000280

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

100103 [病原生物学]; 100218 [急诊医学];

摘要：

Elevated tissue inhibitor of metalloproteinase (TIMP)-1 expression contributes to excess production of extracellular matrix in liver fibrosis. However, there are few Studies on sustained suppression of TIMP-1. We aimed to construct a recombinant adeno-associated virus (AAV) carrying small interfering RNAs (siRNAs) of TIMP-I and investigate the long-term effects of RNA interference upon the TIMP-1 gene in rat hepatic stellate cells (HSCs). Five siRNA oligomers targeting rat TIMP-1 were designed and transfected into HSCs. A U6 promoter followed by the siRNA which had the strongest suppression effect was cloned into the AAV vector and packed into 293 cells to construct the recombinant AAV/ siRNA-TIMP-1/neo. After infecting HSCs with this recombinant AAV, the transcription and expression levels of the TIMP-1 and matrix metalloproteinase-13 (MMP-13) genes were detected at 4 and 12 weeks. Three of the five designed siRNA oligomers had a suppressing effect on TIMP-1 expression in rat HSCs within 72 h. The transcription and expression levels of TIMP-I were suppressed significantly (P<0.05) following recombinant AAV/siRNAI-TIMP-1/neo infection and lasted 12 weeks. TIMP-1 expression in rAAV/ siRNA1-TIMP-1/neo-infected HSCs was Suppressed by 60% after four weeks and 90% after twelve weeks when compared to the control recombinant AAV/neo and uninfected HSCs. Furthermore, the transcription and protein expression levels of MMP-13, the main substrate of TIMP-1, were elevated by similar to 40% at twelve weeks in rAAV/siRNA-TIMP-1/neo-infected HSCs. RNA interference exerts suppressive effect on the TIMP-1 gene in cultured HSCs for a longer time when a recombinant AAV IS utilized as the gene delivery vector.

引用

页码：685 / 692

页数：8

共 24 条

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Comparison of antisense oligonucleotides and siRNAs in cell culture and in vivo [J].