Gene delivery by dendrimers operates via different pathways in different cells, but is enhanced by the presence of caveolin
Gene delivery by dendrimers operates via different pathways in different cells, but is enhanced by the presence of caveolin
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DOI：
10.1016/j.jim.2006.06.007
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发表时间：
2006-07-31
影响因子：
2.2
通讯作者：
George, Andrew J. T.
中科院分区：
文献类型：
--
作者：
Manunta, Maria;Nichols, Benjamin J.;George, Andrew J. T.
为了优化和提高树状大分子介导转染的效果，有必要充分了解这些复合物进入细胞和细胞内运输的机制。以前，我们已经证明了树状大分子的基因传递依赖于胆固醇和膜筏。filipin III对转染的抑制表明，基因传递可能是通过小窝蛋白依赖途径发生的。因此，我们使用表达少量小泡的细胞系（HeLa和HepG2）研究了树突的内化和转染特性。我们表明，与其他细胞相比，胆固醇消耗不会影响树突转染这些细胞的能力。对网格蛋白非依赖性、吞噬和巨噬细胞途径的抑制也未能抑制这些细胞和内皮细胞的转染。然而，过表达caveolin I导致树突突摄取HeLa、HepG2和内皮细胞的速率增加，并提高转染效率。此外，在内皮细胞中，共聚焦显微镜显示树突和小窝蛋白共定位。这些数据强调，树突可能在不同的细胞中使用不同的内化途径，并且小泡形成了非病毒载体传递基因的优先途径。(c) 2006 Elsevier B.V.版权所有
In order to optimise and improve the efficacy of transfection mediated by dendrimers, it is essential to fully understand the mechanisms of cell entry and intracellular trafficking by these complexes. Previously, we have shown that gene delivery by dendrimers is dependent from cholesterol and membrane rafts. The inhibition of transfection by treatment with filipin III suggested that gene delivery might be occurring by a caveolin-dependent pathway. We therefore investigated the internalisation and transfection properties of dendriplexes using cell lines (HeLa and HepG2) that express few caveolae. We show that, in contrast to other cells, cholesterol depletion does not affect the ability of dendriplexes to transfect these cells. Inhibition of clathrin-independent, phagocytic and macropinocytic pathways also failed to inhibit transfection of these cells and endothelial cells. However, overexpression of caveolin I resulted in an increased rate of dendriplex uptake into HeLa, HepG2 and endothelial cells, and increased transfection efficiency. Furthermore, in endothelial cells, confocal microscopy demonstrated colocalisation of dendriplexes and caveolin 1. These data highlight that dendriplexes may use different internalisation pathways in different cells, and that caveolae form a preferential route for gene delivery by this non-viral vector. (c) 2006 Elsevier B.V. All rights reserved.