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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

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【作者】 赵璐陈欢陈丹虎义平余海王青青汤谷平胡秀荣

【Author】 ZHAO Lu1,CHEN Huan1,CHEN Dan1,HU Yi-ping1,YU Hai2,WAN Qing-qing2,TANG Gu-ping1,HU Xiu-rong3 (1.Institute of Chemical Biology and Pharmaceutical Chemistry,Zhejiang University,Hangzhou 310028,China; 2.Institute of Immunology,College of Medicine,Zhejiang University,Hangzhou 310058,China; 3.Center of Analysis and Measurement,Zhejiang University,Hangzhou 310028,China)

【机构】 浙江大学化学生物和药物研究所浙江大学免疫学研究所浙江大学分析测试中心

【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.

【基金】 国家重点基础研究(973)资助项目(2004CB518802);国家高科技研究发展计划(863计划)项目(2007AA03Z355);国家自然科学基金项目(30571068);浙江省自然科学基金重点项目(Z207572)
  • 【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
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