Godbey W T, Wu K K, Mikos A G
Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77251-1892, USA.
Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):5177-81. doi: 10.1073/pnas.96.9.5177.
Poly(ethylenimine) (PEI) is one of a number of polycations that has been used successfully to transfer genes into living cells. Although PEI shows promise in the field of gene therapy, to date no rigorous proof of mechanism has been published regarding the fate of PEI/DNA administered for transfection. Here we show, by using fluorescent labeling and confocal microscopy, the paths of PEI/DNA complexes from endocytosis to gene expression. We found that complexes attach to cell surfaces and migrate into clumps that are endocytosed. The endocytotic vesicles grow in number and size and are occasionally seen to lyse. Most interesting is the fact that endocytosed PEI, whether administered with or without DNA, undergoes nuclear localization in the form of ordered structures.
聚乙烯亚胺(PEI)是众多已成功用于将基因导入活细胞的聚阳离子之一。尽管PEI在基因治疗领域显示出前景,但迄今为止,尚未有关于用于转染的PEI/DNA命运的严格机制证明发表。在这里,我们通过使用荧光标记和共聚焦显微镜,展示了PEI/DNA复合物从内吞作用到基因表达的路径。我们发现复合物附着在细胞表面并迁移到被内吞的团块中。内吞小泡的数量和大小增加,偶尔可见其裂解。最有趣的是,无论是否与DNA一起施用,内吞的PEI都会以有序结构的形式进行核定位。