用于体外基因转染的氨基硅烷Fe3O4复合纳米粒子的合成及表征

高等学校化学学报 ›› 2009, Vol. 30 ›› Issue (12): 2331.

用于体外基因转染的氨基硅烷Fe₃O₄复合纳米粒子的合成及表征

李文章¹, 李洁¹ , 谭泽明²

1. 中南大学化学化工学院, 长沙 410083;
2. 中南大学湘雅医院神经外科, 长沙 410078

收稿日期:2009-03-06 出版日期:2009-12-10 发布日期:2009-12-10
通讯作者: 李洁，女，博士, 教授，博士生导师，从事物理化学及无机材料合成与性能研究. E-mail： lijieliu@mail.csu.edu.cn
基金资助:
教育部新世纪优秀人才资助计划(批准号： NCET-05-0691)和湖南省重大科技计划项目(批准号： 2008SK1001)资助.

Synthesis and Characterization of APTTS-Fe₃O₄ Nanoparticles for in vitro Transfection as Gene Carriers

LI Wen-Zhang¹, LI Jie^1*, TAN Ze-Ming²

1. College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China;
2. Neurosurgery Department of Xiang-Ya Hospital, Central South University, Changsha 410078, China

Received:2009-03-06 Online:2009-12-10 Published:2009-12-10
Contact: LI Jie. E-mail： lijieliu@mail.csu.edu.cn
Supported by:
教育部新世纪优秀人才资助计划(批准号： NCET-05-0691)和湖南省重大科技计划项目(批准号： 2008SK1001)资助.

摘要/Abstract

摘要：

利用2-吡咯烷酮和乙酰丙酮铁为原料制备Fe₃O₄磁性纳米颗粒, 用XRD和TEM对样品进行了表征. 选择偶联剂γ-氨丙基三乙氧基硅烷[NH₂C₃H₆Si(OC₂H₅)₃]对纳米粒子进行表面修饰, 制得APTTS/Fe₃O₄复合载体材料. 以此复合粒子作为传递载体, 将CD基因转染U251胶质瘤细胞. 采用RT-PCR, Western blot及免疫荧光等方法检测CD基因的表达及功能. 结果表明, 制备的Fe₃O₄颗粒粒径为8~10 nm, 结晶度较高; 经表面修饰后, 粒子表面负载—OH, —NH, —NH₂, —C—O和—C—OH等多种功能基团. DNA结合分析及DNase-I消化结果表明, APTTS/Fe₃O₄粒子能够有效地结合和保护DNA. 体外细胞转染实验证实, 该复合纳米颗粒能够高效地传递CD基因进入U251胶质瘤细胞内, 并进行稳定表达.

关键词: 纳米四氧化三铁; 磁性材料; 体外转染; 基因载体

Abstract:

Fe₃O₄ magnetic nanoparticles were prepared via thermal decomposition of Fe(acac)₃ in 2-pyrrolidone. And 3-aminopropyltriethoxy-silane(APTTS) was used to modify the surface of the nanoparticles. The structure and properties of the samples were characterized by means of XRD, TEM and FTIR. The potency of adsorbing DNA and resisting DNase-I digestion of APTTS/Fe₃O₄ were analyzed via agarose gel electrophoresis. Then, APTTS/Fe₃O₄ nanoparticles were evaluated as a kind of plasmid pCMVCD carrier and transferred into human glioma cell line U251 in vitro. The mRNA and protein expression of intracellular CD gene were tested by RT-PCR, Western blot and immunofluorescent staining, respectively. The results show that all the Fe₃O₄ nanoparticles are sized 8—10 nm and have a good crystallinity. Displaying functional group of —OH, —NH, —NH₂, —C—O, —C—OH, the amino-silane modified nanoparticles had the ability of binding DNA and resisting DNase-I digestion. Though the cell transfect test, such composite nanoparticles can transfer DNA into U251 cells efficiently and get stable expression.

Key words: Fe3O4 nanoparticle; Magnetic material; In vitro transfection; Gene carrier

TrendMD:

李文章, 李洁 , 谭泽明. 用于体外基因转染的氨基硅烷Fe₃O₄复合纳米粒子的合成及表征. 高等学校化学学报, 2009, 30(12): 2331.

LI Wen-Zhang, LI Jie*, TAN Ze-Ming. Synthesis and Characterization of APTTS-Fe₃O₄ Nanoparticles for in vitro Transfection as Gene Carriers. Chem. J. Chinese Universities, 2009, 30(12): 2331.

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用于体外基因转染的氨基硅烷Fe₃O₄复合纳米粒子的合成及表征

Synthesis and Characterization of APTTS-Fe₃O₄ Nanoparticles for in vitro Transfection as Gene Carriers

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