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构建络合四氯化铂的PEG-PEI基因载体
Platinum tetrachloride coupled PEG-PEI as drug carrier and gene delivery vector
【摘要】 目的:构建一种能有效抑制肿瘤,且具有一定转染效率的基因载体。方法:在二氯甲烷溶液中将PEG与PEI600偶联,加入四氯化铂的乙醇溶液进行配位反应,生成PEG-PEI-Pt复合物。通过XRD、UV-VIS和FT-IR对配位化合物进行结构表征。凝胶电泳阻滞实验测定PEG-PEI-Pt复合物对DNA的浓缩能力,MTT法测定其在Hela、B16、A293和COS-7细胞上的毒性,体外细胞转染实验测定其在A293和B16细胞上的转染率。结果:XRD、UV-VIS和FT-IR测定结果表明PEI600与PEG的偶联和四氯化铂的配位成功。凝胶电泳组织实验表明PEG-PEI-Pt复合物在与DNA的质量比为0.4∶1时完全阻滞DNA的迁移;MTT结果表明四氯化铂的配位增加了复合物的毒性;体外转染结果表明该复合物的转染率比PEI600高。结论:实验成功构建了络合四氯化铂的PEG-PEI基因新载体。
【Abstract】 Objective:To construct a drug carrier and gene vector PEG-PEI-Pt. Methods: Polyethyleneglycol (PEG) was coupled to polyethylenimine (PEI 600) and platinum tetrachloride; PEG-PEI-Pt complex was formed in ethanol. The complex was characterized by XRD,UV-VIS and FT-IR and the DNA condensation was tested by electrophoretic mobility shift assay. The cell viability was evaluated by MTT assay in Hela,B16,A293 and COS-7 cells and in vitro transfection efficiency was measured in A293 and B16 cells. Results: The structure of PEG-PEI-Pt was characterized by XRD,UV-VIS and FT-IR. PEG-PEI-Pt complex was able to bind DNA at N/P weight ratio of 0.4∶1; the complex showed cytotoxicity on Hela and B16 cells. The complex had higher transfection efficiency in A293 and B16 cells than PEI 600. Conclusion: A novel drug carrier and gene vector PEG-PEI-Pt was constructed successfully.
【Key words】 Polyethyleneimine; Genetic vectors; Transfection; Plasmids; Gene therapy; Cisplatin;
- 【文献出处】 浙江大学学报(医学版) ,Journal of Zhejiang University(Medical Sciences) , 编辑部邮箱 ,2009年01期
- 【分类号】Q78
- 【被引频次】4
- 【下载频次】200