Liver-specific expression of an exogenous gene controlled by human apolipoprotein A-I promoter.
Liver-specific expression of an exogenous gene controlled by human apolipoprotein A-I promoter.
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DOI：
10.1016/j.ijpharm.2010.07.023
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发表时间：
2010-08
影响因子：
5.8
通讯作者：
Yurong Hu;X. Ren;Hui Wang;Yue Ma;Lei Wang;Ying-ying Shen;K. Oka;Zhenzhong Zhang;Yun Zhang
中科院分区：
文献类型：
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作者：
Yurong Hu;X. Ren;Hui Wang;Yue Ma;Lei Wang;Ying-ying Shen;K. Oka;Zhenzhong Zhang;Yun Zhang
肝脏特异性基因治疗有利于减少非靶基因表达可能引起的不良反应。将表达增强型绿色荧光蛋白（EGFP）的质粒pCMV-EGFP的CMV启动子替换为产生pApoAI-EGFP质粒的肝脏特异性启动子载脂蛋白A-I （ApoAI）。在HepG2、SMMC-7721、MCF7、ACC-2和Lo2等多种细胞系中进行的体外表达实验表明，pCMV-EGFP处理在所有细胞系中均能引起基因表达，而pApoAI-EGFP处理仅在肝癌细胞HepG2、SMMC-7721和正常肝细胞Lo2中诱导EGFP表达。pCMV-EGFP或pApoAI-EGFP被配制成聚乙二醇化免疫脂多复合物（PILP），一种新型高效的基因传递系统。在H22荷瘤小鼠静脉注射PILP后，pCMV-EGFP处理小鼠的肝脏、肿瘤、脾脏、脑和肺中均有显著的EGFP表达，而pApoAI-EGFP处理小鼠仅在肝脏和肿瘤中有基因表达，非肝器官基因表达被消除。这项研究表明，使用PILP技术和肝脏特异性启动子可以有效地表达外源基因。
Liver-specific gene therapy is advantageous to minimize the possible adverse effects caused by non-target gene expression. The CMV promoter of the enhanced green fluorescent protein (EGFP) expressing plasmid pCMV-EGFP was replaced with the liver-specific promoter apolipoprotein A-I (ApoAI) generating pApoAI-EGFP plasmid. In vitro expression experiments performed in various cell lines including HepG2, SMMC-7721, MCF7, ACC-2 and Lo2 indicated that pCMV-EGFP treatment caused gene expression in all the cell lines, whereas pApoAI-EGFP treatment only induced EGFP expression in cells of liver origin including the liver cancer cells HepG2 and SMMC-7721 and the normal liver cells Lo2. Either pCMV-EGFP or pApoAI-EGFP was formulated as pegylated immuno-lipopolyplexes (PILP), a novel and efficient gene delivery system. Following intravenous administration of the PILP in H22 tumor-bearing mice, there was significant EGFP expression in liver, tumor, spleen, brain and lung in the pCMV-EGFP treated mice, whereas in the pApoAI-EGFP treated mice there was only gene expression in liver and tumor and the non-liver organ gene expression was eliminated. This study suggests that the use of the PILP technology and liver-specific promoter enables efficient and liver-specific expression of an exogenous gene.