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PEG修饰对PEI聚电解质复合物的药学特性及人乳腺癌细胞的摄取效率影响的研究

刘立婷, 齐宪荣*
  

  1. 北京大学 药学院 药剂学系, 北京 100191
  • 收稿日期:2008-04-25 修回日期:2008-11-10 出版日期:2008-12-15 发布日期:2008-12-15
  • 通讯作者: 齐宪荣*

Effect of poly(ethylene glycol) modified polyethylenimine polyelectrolyte complex on pharmaceutical characteristics and uptake on breast cancer cell

Li-Ting Liu, Xian-Rong Qi*
  

  1. Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China
  • Received:2008-04-25 Revised:2008-11-10 Online:2008-12-15 Published:2008-12-15
  • Contact: Xian-Rong Qi*

摘要: 荧光素阴离子葡聚糖(DFA)和寡核苷酸(ODN)可以和带正电的聚乙亚胺(PEI)通过自组装的方式形成聚电解质复合物。为了调查PEG修饰对PEI聚电解质复合物的药学特性及细胞摄取效率的影响, 我们进行了此项研究。DFAODNPEIPEI-PEG通过吹吸混匀形成聚电解质复合物。我们分别采用动态光散射法及琼脂糖凝胶电泳法对聚电解质复合物的表面性质(粒径, ξ电位), PEIPEI-PEG延滞ODN的能力等进行了评价。MCF-7DFA/PEI DFA/PEI-PEG的摄取效率通过流式细胞术进行测量, 并通过激光扫描聚焦显微镜(CLSM)法来观察DFA/PEIDFA/PEI-PEG聚电解质复合物的细胞内摄作用。 N/P比对ODN/PEI聚电解质复合物的粒径和ξ 电位影响很大, ODN/PEI-PEG复合物的粒径受N/P比的影响要小一些, N/P = 2~16 范围内, ODN/PEI-PEG复合物的粒径在30~100 nm左右。PEIPEI-PEGN/P比等于4时就开始出现延滞ODN的现象, 彻底延滞在N/P比等于8时出现。MCF-7的细胞摄取效率与DFA浓度及转染时间呈正相关, 在适宜条件下, 细胞摄取率可超过99%。本实验结果显示通过隐形修饰的PEI自组装形成的聚电解质复合物可以提高其作为基因载体递送基因进入细胞的能力。

关键词: 聚电解质复合物, 聚电解质复合物, 聚电解质复合物, 聚乙亚胺(PEI), 聚乙亚胺(PEI), 聚乙亚胺(PEI), 细胞摄取率, 细胞摄取率, 细胞摄取率, 隐形修饰, 隐形修饰, 隐形修饰, 乳腺癌细胞, 乳腺癌细胞, 乳腺癌细胞, 自组装, 自组装, 自组装, , ,

Abstract:

Cationic polyethylenimine (PEI) with dextran fluorescein anionic (DFA) or oligodeoxynucleotide (ODN) could form polyelectrolyte complex by self-assembly as a gene delivery vector. This study was designed to investigate the effects on pharmaceutical characteristics and cell uptake PEI after a long-circulation modification with poly(ethylene glycol) (PEG). DFA or ODN reacted with PEI or PEI-PEG to form polyelectrolyte complexes. Surface characters of these complexes and the retardation of ODN by PEI and PEI-PEG were evaluated. The uptake rates of DFA/PEI and DFA/PEI-PEG complexes by MCF-7 cells were evaluated by flow cytometry. Confocal laser scanning microscopy was utilized to visualize the internalization of these complexes. ODN/PEI complex showed the dependence of their size and ξ potential on the N/P ratio. ODN/PEI-PEG complex were much less affected by N/P ratio and their size was around 30 - 100 nm. PEI and PEI-PEG retarded ODN even at N/P ratio as low as 4, and complete retardation was found at N/P ratio of 8. The uptake rate by MCF-7 cells was direct correlated to the DFA concentration and incubation time, and the uptake rate could exceed 99% under the selected condition. The results in this study showed that PEI self-assembly polyelectrolyte complex after stealth or long circulation modification may increase the ability as a gene vector to delivery genes into cells.

Key words: Polyelectrolyte complex, Polyelectrolyte complex, Polyethylenimine, Polyethylenimine, Uptake rate, Uptake rate, Self-assembly, Self-assembly, Breast cancer cell, Breast cancer cell

Supporting: Foundation items: National Nature Science Foundation of China (Grant No. 30772665), Beijing Nature Science Foundation (Grant No. 7083111).
*Corresponding author. Tel.: 86-10-82801584;