Triolein-based polycation lipid nanocarrier for efficient gene delivery: characteristics and mechanism.
Triolein-based polycation lipid nanocarrier for efficient gene delivery: characteristics and mechanism.
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基于三油酸甘油酯的聚阳离子脂质纳米载体用于高效基因递送：特征和机制
DOI：
10.2147/ijn.s24720
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发表时间：
2011
影响因子：
8
通讯作者：
Sha X
中科院分区：
文献类型：
--
作者：
Zhang Z;Fang X;Hao J;Li Y;Sha X
我们建议开发一种具有比我们先前描述的聚阳离子纳米结构脂质纳米载体(PNLC)更高的转染率的聚阳离子脂质纳米载体(PLN)。PLN由三油酸甘油酯、十六烷基低相对分子质量聚乙烯亚胺和二油酰磷脂酰乙醇胺组成。对PLN和PLN/DNA复合体(PDC)的物理化学性质进行了表征。体外转染人肺腺癌(SPC-A1)细胞，并探讨其细胞内机制。测试结果表明，PLN和PDC为均一的带正电荷的纳米粒子。随着甘油三酯含量的增加，pDC的转染率显著提高，且高于pNLC和商品化脂质体™2 0 0 0。特别是，在有10%血清的情况下，PLN的转染率比在无血清的情况下更高。在氯丙嗪和非利平的特异性抑制剂的帮助下，依赖于笼蛋白的内吞途径被确定为PLN介导的SPC-A1细胞成功转染的主要因素。捕获的图像证实，内吞作用后，荧光PDC定位于溶酶体和细胞核中。因此，PLN是一种新型高效的非病毒基因传递载体。
We proposed to develop a polycation lipid nanocarrier (PLN) with higher transfection efficiency than our previously described polycation nanostrucutred lipid nanocarrier (PNLC). PLN was composed of triolein, cetylated low-molecular-weight polyethylenimine, and dioleoyl phosphatidylethanolamine. The physicochemical properties of PLN and the PLN/DNA complexes (PDC) were characterized. The in vitro transfection was performed in human lung adenocarcinoma (SPC-A1) cells, and the intracellular mechanism was investigated as well. The measurements indicated that PLN and PDC are homogenous nanometer-sized particles with a positive charge. The transfection efficiency of PDC significantly increased with the content of triolein and was higher than that of PNLC and commercial Lipofectamine™ 2000. In particular, the transfection of PLN in the presence of 10% serum was more effective than that in its absence. With the help of specific inhibitors of chlorpromazine and filipin, the clathrin-dependent endocytosis pathway was determined to be the main contributor to the successful transfection mediated by PLN in SPC-A1 cells. The captured images verified that the fluorescent PDC was localized in the lysosomes and nuclei after endocytosis. Thus, PLN represents a novel efficient nonviral gene delivery vector.