Lipoplex formulation of superior efficacy exhibits high surface activity and fusogenicity, and readily releases DNA
Lipoplex formulation of superior efficacy exhibits high surface activity and fusogenicity, and readily releases DNA
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DOI：
10.1016/j.bbamem.2006.10.016
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发表时间：
2007-02-01
影响因子：
3.4
通讯作者：
MacDonald, Robert C.
中科院分区：
文献类型：
--
作者：
Koynova, Rumiana;Tarahovsky, Yury S.;MacDonald, Robert C.
已知含有阳离子磷脂二油酰乙基磷脂酰胆碱（EDOPC）和二月桂酰乙基磷脂酰胆碱（EDLPC）的混合物的脂质复合物在转染培养的原代内皮细胞中比单独含有任一脂质的脂质复合物有效得多。大幅度的协同作用允许比较具有非常不同的转染效率但具有微小化学差异的脂质复合物的物理和物理化学性质。在这里，我们报告了EDLPC/EDOPC脂质复合物的上级转染效率与更高的表面活性、与带负电荷的膜模拟脂质体相互作用和混合的更高亲和力以及相对于EDOPC脂质复合物显著更有效的DNA释放相关。对培养细胞的观察结果与模型系统的结果一致：转染的人脐动脉内皮细胞（HUAEC）的共聚焦显微镜显示EDLPC/EDOPC脂质复合物比EDOPC脂质复合物更广泛地将DNA释放到细胞质和核质中;直接固定和包埋在培养皿上的细胞的电子显微镜显示EDLPC/EDOPC脂质复合物与各种细胞膜接触，包括内质网、线粒体和细胞核。概述了有效的脂质转染的事件的顺序进行了讨论的基础上提出的数据。(c)2006 Elsevier B. V.保留所有权利。
Lipoplexes containing a mixture of cationic phospholipids dioleoylethylphosphatidylcholine (EDOPC) and dilauroylethylphosphatidylcholine (EDLPC) are known to be far more efficient agents in transfection of cultured primary endothelial cells than are lipoplexes containing either lipid alone. The large magnitude of the synergy permits comparison of the physical and physico-chemical properties of lipoplexes that have very different transfection efficiencies, but minor chemical differences. Here we report that the superior transfection efficiency of the EDLPC/EDOPC lipoplexes correlates with higher surface activity, higher affinity to interact and mix with negatively charged membrane-mimicking liposomes, and with considerably more efficient DNA release relative to the EDOPC lipoplexes. Observations on cultured cells agree with the results obtained with model systems; confocal microscopy of transfected human umbilical artery endothelial cells (HUAEC) demonstrated more extensive DNA release into the cytoplasm and nucleoplasm for the EDLPC/EDOPC lipoplexes than for EDOPC lipoplexes; electron microscopy of cells fixed and embedded directly on the culture dish revealed contact of EDLPC/EDOPC lipoplexes with various cellular membranes, including those of the endoplasmic reticulum, mitochondria and nucleus. The sequence of events outlining efficient lipofection is discussed based on the presented data. (c) 2006 Elsevier B.V. All rights reserved.