Stimulatory effect of polyethylene glycol (PEG) on gene expression in mouse liver following hydrodynamics-based transfection
Stimulatory effect of polyethylene glycol (PEG) on gene expression in mouse liver following hydrodynamics-based transfection
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DOI：
10.1002/jgm.850
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发表时间：
2006-03-01
影响因子：
3.5
通讯作者：
Kiwada, Hiroshi
中科院分区：
文献类型：
--
作者：
Ishida, Tatsuhiro;Li, Wenhao;Kiwada, Hiroshi
背景快速静脉注射大容量质粒DNA(PDNA)，即基于流体力学的基因转染法，是一种有效的、肝脏特异性的体内基因传递方法。方法采用流体力学方法，研究了在裸露PDNA(荧光素酶)溶液中加入聚乙二醇组分对基因在小鼠肝脏中表达的影响。结果在PDNA溶液中加入1%(w/v)的PEG2000可增强肝脏中基因的表达。将聚乙二醇2000的浓度增加到大于1和高达10%(w/v)时，基因表达水平会降低。相比之下，将聚乙二醇的相对分子质量增加到2000%以上至10000，并不影响基因表达水平。组织病理学和血清化学检测表明，静压或渗透压以浓度依赖的方式增加组织和肝细胞的损伤，并导致基因表达下降。定量评价表明，基因表达的增强是由于稳定了进入肝细胞的pDNA和促进了完整pDNA向细胞核的转运。结论在大多数基因治疗应用和基因功能研究中，导入的基因(S)的持续表达是必要的。这种实现肝脏基因表达增强的简单方法可能在分子和细胞生物学的各种实验室研究中具有巨大的潜力，以及可能在未来的人类临床应用中。版权所有(C)2005 John Wiley&Sons，Ltd.
Background Rapid intravenous injection of a large volume of plasmid DNA (pDNA), i.e. a transfection procedure based on hydrodynamics, is known to be an efficient and liver-specific method of in vivo gene delivery. However, the gene expression is transient.Methods We investigated the effect of addition of polyethylene glycol (PEG) to a solution of naked pDNA (luciferase) on the expression of the gene in mouse liver following transfection by the hydrodynamics-based technique. In addition, the mechanism leading to the enhancement of the gene expression was studied.Results The addition of 1% (w/v) PEG2000 to the pDNA solution enhanced the resulting gene expression in the liver. Increasing the PEG2000 concentration to more than 1 and up to 10% (w/v) rather diminished the gene expression level. By contrast, increasing the molecular weight of PEG to over 2000 up to 10000 did not affect the level of gene expression. Histopathological and serum-chemistry examinations indicated that hydrostatic or osmotic pressure increased tissue and hepatocellular damage in a PEG-concentration-dependent manner, and resulted in a decrease in gene expression. Quantitative evaluation showed that the enhanced gene expression resulted from stabilization of the pDNA introduced into the hepatocytes and an enhancement of the transport of intact pDNA to the nucleus.Conclusions For most gene therapy applications and gene function studies, sustained expression of the introduced gene(s) is necessary. This simple method to achieve enhanced gene expression in liver may have a great potential for a wide variety of laboratory studies in molecular and cellular biology as well as possibly for future clinical applications in humans. Copyright (c) 2005 John Wiley & Sons, Ltd.