Peking University Institute of Cardiovascular Sciences, Peking University Health Science Center, Peking University, Beijing 100083, China.
Mol Pharm. 2010 Feb 1;7(1):267-79. doi: 10.1021/mp900241t.
The GM1/caveolin-1 lipid raft mediated endocytosis mechanism was explored for generation 5 and 7 poly(amidoamine) dendrimer polyplexes employing the Cos-7, 293A, C6, HeLa, KB, and HepG2 cell lines. Expression levels of GM1 and caveolin-1 were measured using dot blot and Western blot, respectively. The level of GM1 in the cell plasma membrane was adjusted by incubation with exogenous GM1 or ganglioside inhibitor PPMP, and the level of CAV-1 was adjusted by upregulation with the adenovirus vector expressed caveolin-1 (AdCav-1). Cholera toxin B subunit was employed as a positive control for uptake in all cases. No evidence was found for a GM1/caveolin-1 lipid raft mediated endocytosis mechanism for the generation 5 and 7 poly(amidoamine) dendrimer polyplexes.
采用 Cos-7、293A、C6、HeLa、KB 和 HepG2 细胞系探索了 GM1/ caveolin-1 脂筏介导的内吞作用机制用于第 5 代和第 7 代聚(酰胺-胺)树枝状聚合物多聚物。使用斑点印迹和 Western blot 分别测量 GM1 和 caveolin-1 的表达水平。通过用外源性 GM1 或神经节苷脂抑制剂 PPMP 孵育来调节细胞质膜中的 GM1 水平,并用表达 caveolin-1 的腺病毒载体(AdCav-1)上调 CAV-1 水平。霍乱毒素 B 亚基被用作所有情况下摄取的阳性对照。没有证据表明第 5 代和第 7 代聚(酰胺-胺)树枝状聚合物多聚物存在 GM1/ caveolin-1 脂筏介导的内吞作用机制。