【摘要】 目的:多肽MC10(MARAKEGGGC)介导的聚乙烯亚胺-β-环糊精聚合物载体作为新型靶向性非病毒基因载体的研究。方法:低分子量的聚乙烯亚胺(PEI)通过羰二咪唑(1,1’-carbonyl-diimidazole,CDI)偶联β-环糊精(β-CyD)合成聚乙烯亚胺-β-环糊精(PEI-β-CyD)作为载体骨架,用琥珀酰亚胺-3-(2-嘧啶二硫)丙酸酯[N-succinimidy-3-(2-pyridyldithio)propionate,SPDP]将MC10偶联到PEI-β-CyD上,制成新型转基因载体材料MC10-PEI-β-CyD。用核磁共振氢谱(1H-NMR)对载体的结构进行化学表征,凝胶电泳阻滞实验研究载体材料对质粒DNA的浓缩能力,透射电镜观察其浓缩质粒DNA后形成的微粒粒径。MTT法检测聚合物的毒性,用SKOV-3、A549和MCF-7细胞株进行体外转基因实验,确定MC10-PEI-β-CyD具有基因携带能力及靶向选择性。结果:成功地合成了MC10-PEI-β-CyD,且MC10-PEI-β-CyD在N/P为5时,能够有效浓缩质粒DNA,形成粒径约为(170±35)nm的微粒。结论:MC10-PEI-β-CyD具有低毒和高转基因效率,对Her-2受体的肿瘤细胞具有很好的靶向性。更多还原

【Abstract】 Objective:To develop a novel non-viral gene delivery vector based on polyethylenimine and β-cyclodextrin targeting to Her-2 receptor (MC10-PEI-β-CyD). Methods: The PEI-β-CyD was synthesized by low molecular weight polyethylenimine (PEI,Mw 600) cross-linked β-cyclodextrin (β-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-β-CyD to form the vector MC10-PEI-β-CyD. The 1H-NMR was used to confirm the structure of vector. The DNA condensing ability,and the particle size of MC10-PEI-β-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. Results: MC10 was linked onto PEI-β-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-β-CyD with low cytotoxicity and high transfection efficiency has been successfully synthesized.更多还原