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

作　　者：赵璐[1] 陈欢[1] 陈丹[1] 虎义平[1] 余海[2] 王青青[2] 汤谷平[1] 胡秀荣[3] 

机构地区：[1]浙江大学化学生物和药物研究所,浙江杭州310028 [2]浙江大学免疫学研究所,浙江杭州310058 [3]浙江大学分析测试中心,浙江杭州310028

出　　处：《浙江大学学报（医学版）》2009年第1期59-66,共8页Journal of Zhejiang University(Medical Sciences)

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)

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。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： Th -PEI-Pt was characterized by XRD,UV-VIS and FT IR. PEC- PEI PI comple DNA at N/P weight ratio of 0.4 ： 1 ; the complex showed cytotoxicity on Hela e structure of PEG x was able to bind 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.

关 键 词：聚乙烯亚胺 遗传载体 转染 质粒 基因疗法 顺铂 

分 类 号：Q78[生物学—分子生物学]