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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2013, Vol. 33 Issue (4): 1-8    
研究报告     
应用改构的炭疽毒素作为靶向肿瘤细胞的药物递送系统及其效果评价
李炳娟1,2, 李玉霞1, 李北平1, 凌焱1, 周围1, 李伟东1, 林海龙1, 梁龙1, 刘刚1, 张景海2, 陈惠鹏1
1. 军事医学科学院生物工程研究所 北京 100071;
2. 沈阳药科大学 沈阳 110016
Construction and Evaluation of the Mutated Anthrax Toxin Proteins as Drug Deliver System for Targeting Tumor Cells
LI Bing-juan1,2, LI Yu-xia1, LI Bei-ping1, LING Yan1, ZHOU Wei1, LI Wei-dong1, LIN Hai-long1, LIANG Long1, LIU Gang1, ZHANG Jin-hai2, CHEN Hui-peng1
1. Beijing Institute of Biotechnology, Beijing 100071, China;
2. Department of Life Science, ShenYang Pharmaceutical University, ShenYang 110016, China
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摘要: 目的:通过改造炭疽毒素保护性抗原Protective Antigen (PA)及致死因子Lethal Factor (LF),尝试建立更加广谱的新型炭疽毒素靶向给药系统并对其递送效率进行定量评价。方法:采用基因工程手段,分别构建了3种改构的天然炭疽毒素保护性抗原PA及炭疽毒素的LF N端融合海肾荧光素酶(Luciferase)的LFn-linker-Luc的大肠杆菌重组表达体系。利用CCK-8法评价改构PA和LF共同作用肿瘤细胞后的细胞存活率;利用改构PA和LFn-linker-Luc与肿瘤细胞共孵育,通过测定细胞内荧光素酶活性,评价改构PA靶向肿瘤细胞的效果。结果:体外酶解实验证明构建的改构PA蛋白能够被正确地酶解成目的大小的片段;改构PA和LF共同作用肿瘤细胞能够显著降低细胞存活率;利用LFn-linker-Luc能够评价改构PA的靶向效率,PA蛋白的改构方式与其递送效率相关。结论:设计并改构的炭疽毒素药物递送系统,能够实现特异性靶向肿瘤细胞的效果,并具有更广谱的作用效果,为研制新型广谱抗肿瘤药物提供了新的思路和方法。
关键词: 炭疽毒素保护性抗原荧光素酶基质金属蛋白酶肿瘤靶向治疗    
Abstract: Objective: The anthrax toxin proteins constitute a unique membrane translocation system which provide an attractive model system for the development of reagents for cell biology and therapeutics. The mutated anthrax toxin proteins PA and LF were constructed to target to tumor cells. Methods: The recombinant Rosseta(DE3) which harboring three kinds of mutated PA and LF were constructed separately. The cell viability after incubated with mutated PA and LF was detected by cytotoxicity assay. The ability of the mutated PA translocation system targeting to tumor cells was measured by the amount of the luciferase of intra-cellular. Results: The mutated PA can be cleaved by matrix metalloproteinases or uPA in vitro correctly. When the tumor cell incubated with the mutated PA and LF,the cell viability was decreased remarkably. The amount of the luciferase of intra-cellular can correctly reflex the ability of the mutated PA translocation system targeting to tumor cells. Conclusions: These mutated PA proteins showed natural biological activities and had the ability of targeting to tumor cells. These results provide a new insight for the study of the antitumor drugs.
Key words: Anthrax toxin    Protective antigen    Luciferase    Matrix metalloproteinases    Targeting tumor therapy
收稿日期: 2012-08-14 出版日期: 2013-04-25
ZTFLH:  Q819  
基金资助: 国家自然科学基金(81000763);国家科技重大专项(2012ZX09301003);国家"973"课题(2009CB52604)资助项目
通讯作者: 凌焱, 陈惠鹏     E-mail: chenhp0909@163.com;lingny@me.com
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张景海
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引用本文:

李炳娟, 李玉霞, 李北平, 凌焱, 周围, 李伟东, 林海龙, 梁龙, 刘刚, 张景海, 陈惠鹏. 应用改构的炭疽毒素作为靶向肿瘤细胞的药物递送系统及其效果评价[J]. 中国生物工程杂志, 2013, 33(4): 1-8.

LI Bing-juan, LI Yu-xia, LI Bei-ping, LING Yan, ZHOU Wei, LI Wei-dong, LIN Hai-long, LIANG Long, LIU Gang, ZHANG Jin-hai, CHEN Hui-peng. Construction and Evaluation of the Mutated Anthrax Toxin Proteins as Drug Deliver System for Targeting Tumor Cells. China Biotechnology, 2013, 33(4): 1-8.

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https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I4/1

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