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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2015, Vol. 35 Issue (1): 54-60    DOI: 10.13523/j.cb.20150108
技术与方法     
适配体介导脂质体靶向递送siRNA的研究
唐德平1, 毛爱红2, 王芳2, 张虹2, 王黎2, 廖世奇2
1. 兰州交通大学化学与生物工程学院 兰州 730070;
2. 甘肃省医学科学研究院 兰州 730050
Targeted Delivery of siRNA Mediated by Aptamer Modified Liposome
TANG De-ping1, MAO Ai-hong2, WANG Fang2, ZHANG Hong2, WANG Li2, LIAO Shi-qi2
1. The School of Chemical & Biological Engineering, Lanzhou Jiaotong University, Lanzhou 730070, China;
2. Institute of Gansu Medical Science Research, Lanzhou 730050, China
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摘要：

为探讨适配体介导的脂质体靶向递送siRNA的可行性,采用前列腺癌细胞膜表面抗原(PSMA)的适配体A10-3.2与脂质体结合,构建适配体-脂质体靶向递送体系(Apt -LP),并利用Apt-LP递送pEGFP-N1质粒和Bcl2 siRNA到前列腺癌细胞LNCaP(PSMA+)和PC-3(PSMA-),转染48h后,用荧光显微镜检测绿色荧光蛋白的表达,qPCR检测 Bcl2 mRNA表达,蛋白印记法检测Bcl2蛋白表达,Hoechst 33258核染法分析体外抗肿瘤活性。结果显示:与脂质体递送体系相比,Apt-LP显著提高递送pEGFP-N1质粒和Bcl2 siRNA到靶细胞LNCaP(PSMA+)的效率;显著提高Bcl2 siRNA 诱导的靶细胞LNCaP(PSMA+) Bcl2 基因沉默效应,更有效的诱导靶细胞LNCaP(PSMA+)凋亡。结果表明:Apt -LP是一种有效的siRNA靶向递送体系,具有潜在的临床应用价值。
关键词： siRNA;  脂质体;  靶向递送;  适配体    
Abstract:

To investigate the efficiency of siRNA using aptamer-conjugated liposome(Apt-LP),A10-3.2 was used as a PSMA-targeting ligand in the design of a liposome-based siRNA delivery system to prostate cancer cells. The sequence of the aptamer used in this study is modified with 3'- Cholesterol and with 2'- Fluor pyrimidines. The cholesterol tag immobilizes the aptamer on the liposome surface by inserting into the hydrophobic lipid membrane. Liposome was conjugated to aptamer and used as a vehicle for siRNA target delivery. GFP expression assays of pEGFP - N1 against LNCaP (PSMA+) and PC-3 (PSMA-) cells demonstrated that the transfection efficiency of the synthesized Apt-LP complex was higher than that of the liposome. In addition, gene knock down assay of Apt-LP-siRNA complex in LNCaP (PSMA+) and PC-3 (PSMA-) cells showed that Bcl2 gene silencing effect significantly higher than that of LP-siRNA complex and more effectively induced apoptosis of LNCaP cells. These results suggest that the Apt - LP is a kind of effective siRNA targeting delivery system, has potential clinical application.
Key words: Aptamer    siRNA    Liposome    Targeted delivery
收稿日期: 2014-06-27 出版日期: 2015-01-25
ZTFLH:  Q522  
基金资助:

甘肃省中青年基金(1107RJYA033),兰州交通大学校青年基金(2013012)资助项目
通讯作者: 毛爱红     E-mail: maoaih@aliyun.com
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引用本文:

唐德平, 毛爱红, 王芳, 张虹, 王黎, 廖世奇. 适配体介导脂质体靶向递送siRNA的研究[J]. 中国生物工程杂志, 2015, 35(1): 54-60.

TANG De-ping, MAO Ai-hong, WANG Fang, ZHANG Hong, WANG Li, LIAO Shi-qi. Targeted Delivery of siRNA Mediated by Aptamer Modified Liposome. China Biotechnology, 2015, 35(1): 54-60.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150108    或    https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I1/54


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