Opposing roles of syndecan-1 and syndecan-2 in polyethyleneimine-mediated gene delivery
Opposing roles of syndecan-1 and syndecan-2 in polyethyleneimine-mediated gene delivery
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DOI：
10.1074/jbc.m705424200
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发表时间：
2008-03-21
影响因子：
4.8
通讯作者：
Durocher, Yves
中科院分区：
文献类型：
--
作者：
Paris, Sebastien;Burlacu, Alina;Durocher, Yves
聚乙烯亚胺（PEI）是用于基因转移的有效非病毒载体。硫酸乙酰肝素蛋白聚糖被认为是PEI.DNA复合物（聚合复合物）的细胞表面受体。在这里，我们研究了syndecan-1（SDC 1）和syndecan-2（SDC 2）是否参与PEI介导的转染。向HEK 293细胞添加聚合物后，绿色荧光蛋白标记的SDC迅速与PEI形成簇，这取决于脂筏完整性。然而，虽然SDC 1过表达略有增强PEI介导的基因表达，SDC 2显着抑制it. Confocal显微镜分析表明，SDC1.polyplex内吞作用发生在几分钟内加入polyplexes后，而SDC2.polyplex内吞作用需要几个小时。SDC 1胞质缺失突变体的表达表明，SDC 1胞质尾区是基因表达所需的，但不是聚类或内吞作用，而SDC 1/SDC 2嵌合体的过表达表明，SDC 2胞外域是负责对基因转移的抑制作用。这项研究提供的证据表明，SDC可能对PEI介导的转染具有相反的作用。
Polyethyleneimines (PEIs) are efficient non-viral vectors for gene transfer. Heparan sulfate proteoglycans have been proposed to be the cell-surface receptors for PEI.DNA complexes (polyplexes). Here, we investigated if syndecan-1 (SDC1) and syndecan-2 (SDC2) are involved in PEI-mediated transfection. Following addition of polyplexes to HEK293 cells, green fluorescent protein-tagged SDCs rapidly formed clusters with PEI that were dependent of lipid raft integrity. However, although SDC1 overexpression slightly enhanced PEI-mediated gene expression, SDC2 dramatically inhibited it. Confocal microscopy analysis showed that SDC1.polyplex endocytosis occurred within minutes after addition of polyplexes, whereas SDC2.polyplex endocytosis took hours. Expression of SDC1 cytoplasmic deletion mutants revealed that the SDC1 cytoplasmic tail is required for gene expression, but not for clustering or endocytosis, whereas overexpression of SDC1/SDC2 chimeras showed that the SDC2 ectodomain is responsible for the inhibitory effect on gene transfer. This study provides evidence that SDCs may have opposing effects on PEI-mediated transfection.