Efficient expression of vascular endothelial growth factor using minicircle DNA for angiogenic gene therapy
Efficient expression of vascular endothelial growth factor using minicircle DNA for angiogenic gene therapy
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DOI：
10.1016/j.jconrel.2007.10.014
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发表时间：
2008-01-22
影响因子：
10.8
通讯作者：
Kim, Sung Wan
中科院分区：
文献类型：
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作者：
Chang, Chien-Wen;Christensen, Lane V.;Kim, Sung Wan
基于质粒DNA（pDNA）的基因治疗的应用受到其低效的转基因表达的限制。在这项研究中，小环DNA的有效血管内皮生长因子（VEGF）在骨骼肌细胞的表达进行了评估。通过半定量电泳方法研究编码VEGF的小环DNA的产生，从而优化细菌培养条件并产生高纯度（86.6%）的小环DNA。在SV 40启动子控制下的VEGF小环DNA（pMmi-SV-VEGF）在两种不同的骨骼肌细胞系（C2 C12和L 8）中显示出VEGF mRNA的量增加，并且VEGF表达比常规质粒（pSV-VEGF）高多达8倍。在C2 C12骨骼肌细胞系中测试具有不同启动子（包括SV 40、CMV和鸡P-肌动蛋白）的小环DNA的VEGF表达。微环DNA产生的高VEGF表达在体外刺激有效的内皮细胞生长。此外，与常规质粒相比，小环DNA在小鼠胫骨前肌（TA）中表达更高的VEGF。两者合计，结果表明，微环DNA是一个有效的基因载体血管生成VEGF表达在骨骼肌，并可能是有用的治疗外周动脉疾病（PAD）。(C)2007 Elsevier B. V.保留所有权利。
The application of plasmid DNA (pDNA)-based gene therapy is limited by its inefficient transgene expression. In this study, minicircle DNA was evaluated for efficient vascular endothelial growth factor (VEGF) expression in skeletal muscle cells. Production of minicircle DNA encoding VEGF was studied by a semi-quantitative electrophoresis method leading to optimized bacterial culture conditions and producing high purity (86.6%) minicircle DNA. The VEGF minicircle DNA under control of the SV40 promoter (pMmi-SV-VEGF) showed an increased amount of VEGF mRNA and up to 8 times more VEGF expression than a conventional plasmid (pSV-VEGF) in two different skeletal muscle cell lines (C2C12 and L8). Minicircle DNA with different promoters, including the SV40, CMV and chicken P-actin, was tested for VEGF expression in a C2C12 skeletal muscle cell line. The high VEGF expression generated by minicircle DNA stimulated efficient endothelial cell growth in vitro. Furthermore, minicircle DNA expressed higher VEGF compared to conventional plasmid in the tibialis anterior (TA) muscle of mice. Taken together, the results suggest that minicircle DNA is an efficacious gene vector for angiogenic VEGF expression in skeletal muscle and may be useful for treating peripheral arterial disease (PAD). (C) 2007 Elsevier B.V. All rights reserved.