[肽MC10介导的聚乙烯亚胺-β-环糊精作为靶向人表皮生长因子受体2的基因递送载体]
[Peptide MC10 mediated PEI-beta-CyD as a gene delivery vector targeting to Her-2 receptor].
作者信息
Liu Jun, Hu Yi-Ping, Jiang Qi-Ying, Chen Dan, Yu Hai, Wang Qing-Qing, Tang Gu-Ping
机构信息
Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou 310028, China.
出版信息
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2009 Jan;38(1):7-14. doi: 10.3785/j.issn.1008-9292.2009.01.002.
OBJECTIVE
To develop a novel non-viral gene delivery vector based on polyethylenimine and beta-cyclodextrin targeting to Her-2 receptor (MC10-PEI-beta-CyD).
METHODS
The PEI-beta-CyD was synthesized by low molecular weight polyethylenimine (PEI, Mw 600) cross-linked beta-cyclodextrin (beta-CyD) via N, N-carbonyldiimidazole (CDI). The chemical linker[N-succinimidy-3-(2-pyridyldithio) propionate, SPDP] was used to bind peptide MC10 (MARAKEGGGC) to PEI-beta-CyD to form the vector MC10-PEI-beta-CyD. The (1)H-NMR was used to confirm the structure of vector. The DNA condensing ability,and the particle size of MC10-PEI-beta-CyD/DNA complexes were demonstrated by gel retardation assay and electron microscope observation (TEM). Cell viability was tested by MTT assay. The transfection efficiency was determined on cultured SKOV-3, A549 and MCF-7 cells.
RESULT
MC10 was linked onto PEI-beta-CyD successfully. The vector was able to condense DNA at N/P ratio of 5 and particle size was about (170 +/-35)nm. The vector showed low cytotoxicity and high transfection efficiency in cultured SKOV-3, A549 and MCF-7 cells.
CONCLUSION
A novel non-viral vector MC10-PEI-beta-CyD with low cytotoxicity and high transfection efficiency has been successfully synthesized.
目的
基于聚乙烯亚胺和β-环糊精开发一种靶向人表皮生长因子受体2（Her-2）的新型非病毒基因递送载体（MC10-PEI-β-CyD）。
方法
通过低分子量聚乙烯亚胺（PEI，Mw 600）经N，N-羰基二咪唑（CDI）交联β-环糊精（β-CyD）合成PEI-β-CyD。使用化学连接剂[N-琥珀酰亚胺基-3-（2-吡啶基二硫代）丙酸酯，SPDP]将肽MC10（MARAKEGGGC）与PEI-β-CyD结合以形成载体MC10-PEI-β-CyD。采用氢核磁共振波谱（1H-NMR）确认载体结构。通过凝胶阻滞试验和电子显微镜观察（TEM）证明MC10-PEI-β-CyD/DNA复合物的DNA凝聚能力和粒径。采用MTT法检测细胞活力。在培养的SKOV-3、A549和MCF-7细胞上测定转染效率。
结果
MC10成功连接到PEI-β-CyD上。该载体在N/P比为5时能够凝聚DNA，粒径约为（170±35）nm。该载体在培养的SKOV-3、A549和MCF-7细胞中显示出低细胞毒性和高转染效率。
结论
成功合成了一种具有低细胞毒性和高转染效率的新型非病毒载体MC10-PEI-β-CyD。
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