| 王玉强1 臧怡2 蔡文玮1 陈谊1 金拓2 苏靖2△ 盛净1△.交联聚乙烯亚胺衍生物的构建及其转染COS-7 细胞的研究[J].,2012,12(10):1805-1807 |
| 交联聚乙烯亚胺衍生物的构建及其转染COS-7 细胞的研究 |
| Construction of Cross-linked Polyethylenimine Derivative and ItsTransfection in COS-7 Cells |
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| DOI： |
| 中文关键词: 聚乙烯亚胺 非病毒基因载体 转染效率 细胞毒性 |
| 英文关键词: Polyethylenimine Non-viral gene carrier Transfection efficiency Cytotoxicity |
| 基金项目:国家自然科学基金资助项目（81001416），上海市科委基金项目（10JC1408902） |
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| 中文摘要: |
| 目的：优化构建交联聚乙烯亚胺（Polyethylenemine,PEI）衍生物PEI-Bu，研究其对非洲绿猴肾成纤维细胞系（COS-7）的转染
活性和细胞毒性。方法：以PEI 800Da 为骨架，1,4- 丁二醇二氯甲酸酯为连接剂制备聚合物PEI-Bu，琼脂糖凝胶电泳考察其复合
质粒DNA 的能力，MTT 法检测PEI-Bu 对COS-7 的毒性，以荧光素酶质粒作为报告基因，测定PEI-Bu/DNA 复合物在COS-7 细
胞的转染活性。结果：凝胶电泳表明PEI-Bu/DNA 在质量比大于1 时即具有复合DNA 的能力，PEI-Bu 的细胞毒性随浓度增大而
增大，在同一浓度下PEI-Bu 的细胞毒性小于PEI 25kDa,(P<0.05), PEI-Bu/DNA 在质量比为5 时达到最高转染活性，高于PEI 25
kDa(P<0.01)，并与Lipofectamine2000 相当（P>0.05）。结论：PEI-Bu 在COS-7 细胞中是一种低细胞毒性、高转染活性的非病毒基
因载体(与商业化的PEI 25kDa 比较)，其在基因治疗领域中具有潜在的应用前景。 |
| 英文摘要: |
| Objective: To synthesize cross-linked Polyethylenimine derivative with modified procedure and investigate its
cytotoxicity and transfection efficiency on COS-7 cells. Methods: PEI-Bu was synthesized by using PEI 800Da as a backbone and 1,
4-Dibutanediol Chloroformate was used as a linker. The pDNA condensation ability of the polymer was evaluated by agarose gel
electrophoresis. MTT assay was employed to measure the cytotoxicity of the polymer. Luciferase plasmid was used as the reporter gene
to investigate transfection efficiency in COS-7 cells. Results: Gel retardation assay showed complete condensation of pDNA at w/w ratios
exceeded 1. Cytotoxicity of PEI-Bu increased with the increasing of the concentrations. PEI-Bu showed lower cytotoxicity than PEI
25kDa at the same concentration. Transfection result indicated that the polymer performed the highest transfection efficiency at w/w 5,
which was higher than that in commercially available PEI 25kDa (P<0.01) and comparable with Lipofectamine 2000（P>0.05）.
Conclusion: The polymer PEI-Bu displayed much lower cytotoxicity and significantly enhanced transfection efficiency than PEI 25kDa in
COS-7 cells, Therefore, it would be a promising candidate for safe and efficient gene delivery in gene therapy. |
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