Dubruel Peter, Christiaens Bart, Rosseneu Maryvonne, Vandekerckhove Joël, Grooten Johan, Goossens Vera, Schacht Etienne
Polymer Materials Research Group, Department of Organic Chemistry, Ghent University, Ghent, Belgium.
Biomacromolecules. 2004 Mar-Apr;5(2):379-88. doi: 10.1021/bm034438d.
Recently, we have shown that polymethacrylates containing imidazole side groups (HYMIMMA) or acid functions (MA), which have similar buffering properties as polyethyleneimine, were not able to transfect Cos-1 cells, whereas polymers containing only tertiary amines (DMAEMA) do transfect Cos-1 cells (Dubruel, P. et al. Eur. J. Pharm. Sci. 2003, 18 (3-4), 211-220). In the present work, we investigated to what extent the differences in transfection activity are related to differences in cellular internalization and/or subcellular localization. Therefore, we synthesized a series of polymethacrylates containing primary amine functions, used for the coupling of the fluorescent Oregon Green probe. The polymers containing acid functions were labeled with an amine containing fluorescein derivative (5-aminomethyl)fluorescein hydrochloride. It is demonstrated that the endosomal release of the MA and HYMIMMA-based complexes might be the limiting step in the gene transfer process in Cos-1 cells.
最近，我们已经表明，含有咪唑侧基（HYMIMMA）或酸性官能团（MA）的聚甲基丙烯酸酯，其具有与聚乙烯亚胺相似的缓冲性能，但不能转染Cos-1细胞，而仅含叔胺的聚合物（DMAEMA）则能够转染Cos-1细胞（Dubruel，P.等人，《欧洲药理学杂志》，2003年，18（3 - 4），211 - 220）。在本研究中，我们调查了转染活性的差异在多大程度上与细胞内化和/或亚细胞定位的差异相关。因此，我们合成了一系列含有伯胺官能团的聚甲基丙烯酸酯，用于偶联荧光俄勒冈绿探针。含有酸性官能团的聚合物用含胺的荧光素衍生物（5 - 氨基甲基）荧光素盐酸盐进行标记。结果表明，基于MA和HYMIMMA的复合物从内体释放可能是Cos-1细胞基因转移过程中的限制步骤。
