携siRNA真核表达载体靶向脂质微泡的构建及其体外靶向性研究  

Preparation of Microbubble Carrying Eukaryotic Expression Vector of siRNA and Herceptin and its Targeting Ability in Vitro

作　　者：魏巍丽[1] 林敏[1] 陈志奎[1] 林礼务[1] 薛恩生[1] 何以敉[1] 俞丽云[1] 

机构地区：[1]福建医科大学附属协和医院超声科,福州市350001

出　　处：《中国超声医学杂志》2014年第5期454-456,共3页Chinese Journal of Ultrasound in Medicine

基　　金：福建省卫生厅医学创新课题(No.2012-CXB-15)

摘　　要：目的构建携siRNA真核表达载体的靶向脂质微泡,评估其体外靶向乳腺癌细胞能力。方法合成聚乙烯亚胺胆固醇PEI-Chol,观察其对质粒DNA的包载与保护作用;反相挥发-机械振荡法制备携siRNA真核表达载体的脂质微泡,以生物素-亲和素桥接法将抗HER2抗体Herceptin连接到微泡,观察微泡形态、粒径及荧光;观察微泡与乳腺癌细胞SK-BR-3特异性结合能力。结果合成的PEI-Chol在N/P比为3.75时可完全包裹质粒DNA,并保护DNA免受核酸酶降解;制备的微泡平均粒径(3.8±0.9)μm,表面发出红色、绿色荧光,并能与乳腺癌细胞特异性结合。结论携siRNA真核表达载体靶向脂质微泡可特异结合乳腺癌细胞,有望为乳腺癌靶向治疗研究提供一种基因转染新方法。Objective To construct targeted microbubbles carrying eukaryotic expression vector of siRNA,and explore its targeting ability in vitro.Methods Polyethyleneimine cholesterol （PEI-Chol） was synthesized chemically,and its abilities of carrying and protecting plasmid DNA were observed as well.The microbubbles carrying eukaryotic expression vector were prepared by the method of inverting volatile mechanical oscillation,Herceptin was connected to the microbubbles with Biotin-Avidin bridging method,and the form,particle size,fluorescence and ability of specific binding to breast cancer cells were observed.Results Plasmid DNA can be completely wrapped by PEI-Chol and protected from nuclease degradation at an N/P ratio of 3.75.The prepared microbubbles were spherical in shape with an average particle size of （3.8±0.9）μm,emitted red,green fluorescence on the surface,and can bind to SK-BR-3 cells specifically.Conclusions The targeted microbubbles carrying eukaryotic expression vector of siRNA can specifically bind to breast cancer cells,which would provide an effective method for gene transfection.

关 键 词：超声造影剂 乳腺癌 真核表达载体 HERCEPTIN 

分 类 号：R445.1[医药卫生—影像医学与核医学] R737.9[医药卫生—诊断学]