摘要
目的合成对叔丁基杯[4]芳烃酸偶合低相对分子质量聚乙烯亚胺聚合物作为非病毒转基因载体,并评价其毒性、压缩DNA的能力,携带报告基因转染细胞的能力。方法采用新方法合成对叔丁基杯[4]芳烃酸偶合聚乙烯亚胺聚合物,通过核磁共振方法表征。将合成的新聚合物与DNA混合,得到新聚合物/DNA复合物,用琼脂糖电泳试验测定不同N/P比值形成复合物时对质粒DNA电泳的阻滞情况,评价其压缩DNA能力。透射电镜法检测复合物的大小,MTT法检测其对细胞的毒性作用,使用新聚合物携带报告基因转染细胞,并与PEI25000比较转染率。结果新聚合物压缩质粒DNA的能力随N/P比值增大而增强,在N/P比为6时可以完全阻滞质粒DNA的电泳,在N/P比为60时,复合物粒径约291 nm,复合物的表面电荷约14.6 m V。细胞毒性试验表明新聚合物与PEI25000相比毒性明显下降,携带报告基因转染MCF-7细胞转染效率与PEI25000相近。结论对叔丁基杯[4]芳烃酸聚乙烯亚胺新聚合物具有较强压缩质粒DNA能力,对细胞毒性低,转染效率高,是一种可应用于基因治疗的新型非病毒载体。
Objective To study the synthesis and function of p-tert-butyl-Calix（ 4） arene acid coupled to low molecular weight polyethylenimine as a gene delivery vector.Methods The polymer of p-tert-butyl-Calix（ 4）arene acid coupled to polyethylenimine was synthesized and characterized by NMR. Agarose electrophoresis was used to determine the capacity of polymer / DNA complexes for condensed DNA at different N / P ratios.The size of polymer / DNA complexes was observed by transmission electron microscope. The cytotoxocity in vitro was measured by MTT assay. The capacity of report gene delivery was determined in comparison with PEI25000. Results The capacity of condensed DNA with complexes enhanced with the increasing N / P ratio,and the plasmid DNA was completely retarded when polymer at 6 of N / P ratio. The particle size of complexes was about 291 nm at 60 of N / P ratio. Zeta potential of complexes was about ＋ 14.6 m V at 60 of N / P ratio. The cytotoxicity of the polymer was lower than PEI25000. The gene delivery efficiency of polymer was similar to PEI25000. Conclusion The novel polymer of p-tert-butyl-Calix（ 4） arene coupled to polyethylenimine is a potential non-viral vector in gene therapy with the capacity of condensing DNA,lower cytotoxicity and high gene delivery efficiency.
出处
《广东药学院学报》
CAS
2015年第1期104-109,共6页
Academic Journal of Guangdong College of Pharmacy
基金
广东省教育厅优秀人才培育基金(2050205F5504)
关键词
非病毒载体
芳烃酸
报告基因
转染
non-viral vector
p-tert-butyl-Calix（4） arene acid
report gene
gene delivery
