Efficient gene transfer by pullulan-spermine occurs through both clathrin- and raft/caveolae-dependent mechanisms
Efficient gene transfer by pullulan-spermine occurs through both clathrin- and raft/caveolae-dependent mechanisms
复制标题
DOI：
10.1016/j.jconrel.2006.09.001
复制
发表时间：
2006-11-10
影响因子：
10.8
通讯作者：
Tabata, Yasuhiko
中科院分区：
文献类型：
--
作者：
Kanatani, Isao;Ikai, Tomonori;Tabata, Yasuhiko
探讨了阳离子多糖普鲁兰-精胺作为基因载体的可行性及其作用机制。普鲁兰精胺介导的质粒DNA转染导致细胞毒性大大降低，基因表达水平增加了10倍，相比Lipofectamine 2000，市售的阳离子脂质。此外，用普鲁兰精胺转染p53表达质粒DNA后，T24细胞的体外增殖显著降低，而用Lipofectamine 2000转染则无此作用。普鲁兰-精胺介导的基因表达被网格蛋白介导的内吞抑制剂氯丙嗪和筏/caveolac抑制剂甲基-β-环糊精和菲律宾肽抑制。我们得出结论，普鲁兰精胺是一个很有前途的载体基因转染，普鲁兰精胺质粒DNA复合物的细胞摄取介导的网格蛋白和筏/小窝依赖的内吞途径。(c)2006 Elsevier B. V.保留所有权利。
The feasibility and mechanism of gene delivery by pullulan-spermine, a recently developed cationic polysaccharide, were investigated. Pullulan-spermine-mediated transfection of plasmid DNA resulted in greatly reduced cytotoxicity and a 10-fold increase in the level of gene expression when compared to Lipofectamine 2000, a commercially available cationic lipid. Additionally, after transfection of p53-expressing plasmid DNA by pullulan-spermine but not Lipofectamine 2000, the in vitro proliferation of T24 cells was significantly reduced. Pullulan-spermine-mediated gene expression was inhibited by both chlorpromazine of clathrin-mediated endocytosis inhibitor and methyl-beta-cyclodextrin and filipin of raft/caveolac inhibitors. We conclude that pullulan-spermine is a promising carrier for gene transfection, and that cellular uptake of pullulan-spermine-plasmid DNA complexes is mediated by clathrin- and raft/caveolae-dependent endocytotic pathways. (c) 2006 Elsevier B.V. All rights reserved.