Lampela Pasi, Soininen Päivi, Puttonen Katja A, Ruponen Marika, Urtti Arto, Männistö Pekka T, Raasmaja Atso
Department of Pharmacology and Toxicology, University of Kuopio, PO Box 1627, FIN-70211 Kuopio, Finland.
Int J Pharm. 2004 Oct 13;284(1-2):43-52. doi: 10.1016/j.ijpharm.2004.07.023.
Glycosaminoglycans (GAGs) are negatively charged polysaccharides that are found, e.g. on cell surface. GAGs have been reported to influence gene transfection. We have previously reported that cationic lipid-mediated gene transfection can be improved by combining a small polyethylenimine (PEI) with cationic lipids. In the present study, we examined if GAGs have any effect on the synergism of small PEIs and other cationic carriers. We used wild-type CHO (GAG+) and pgsB-618 cells (GAG-). Transfection efficiency was studied using lacZ and GFP reporter genes. We found that GAGs decreased the overall level of transgene expression in a reagent-dependent manner, but the synergism caused by low-MW PEIs was less affected. There were no major differences between cell lines in cellular uptake or intracellular localization of plasmids when measured with flow cytometry and confocal microscopy, respectively. In conclusion, cell-surface GAGs interfere with transfection efficiency of different cationic reagents, but that is not necessarily related to the synergy of small PEIs and cationic lipids.
糖胺聚糖(GAGs)是带负电荷的多糖,例如存在于细胞表面。据报道,GAGs会影响基因转染。我们之前曾报道,通过将小分子量聚乙烯亚胺(PEI)与阳离子脂质相结合,可以提高阳离子脂质介导的基因转染效率。在本研究中,我们研究了GAGs是否对小分子量PEI与其他阳离子载体的协同作用有任何影响。我们使用了野生型CHO(GAG+)和pgsB - 618细胞(GAG-)。使用lacZ和GFP报告基因研究转染效率。我们发现,GAGs以试剂依赖的方式降低了转基因表达的总体水平,但低分子量PEI引起的协同作用受影响较小。当分别用流式细胞术和共聚焦显微镜测量时,细胞系在质粒的细胞摄取或细胞内定位方面没有重大差异。总之,细胞表面的GAGs会干扰不同阳离子试剂的转染效率,但这不一定与小分子量PEI和阳离子脂质的协同作用相关。