Receptor-mediated gene delivery by folic acid-modified stearic acid-grafted chitosan micelles.
Receptor-mediated gene delivery by folic acid-modified stearic acid-grafted chitosan micelles.
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通过叶酸修饰的硬脂酸接枝壳聚糖胶束进行受体介导的基因递送
DOI：
10.2147/ijn.s23828
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发表时间：
2011
影响因子：
8
通讯作者：
Hu FQ
中科院分区：
文献类型：
--
作者：
Du YZ;Cai LL;Li J;Zhao MD;Chen FY;Yuan H;Hu FQ
背景与病毒载体不同，阳离子聚合物已被认为是低免疫原性基因传递的有效非病毒载体。然而，缺乏器官或细胞的特异性有时会阻碍它们的应用，而聚合物载体的修饰也显示出在实现细胞特异性靶向传递和提高细胞内基因转移效率方面的成功改进。方法通过1-乙基-3-(3-二甲氨基丙基)碳二亚胺偶联反应合成叶酸偶联硬脂酸接枝壳聚糖(FA-CS-SA)胶束，用于特异性受体介导的基因传递。结果利用壳聚糖的阳离子性质，胶束能使载体DNA(PDNA)紧密结合，形成胶束/PDNA复合纳米粒。不同N/P比的FA-CS-SA/DNA复合体的粒径为10 0~2 0 0 nm，−为2 0~−10 mV。DNase I保护实验表明，这些化合物能有效地保护缩合DNA免受DNase I的酶降解。细胞毒性实验表明，与LipofeTamineTM2000相比，这些胶束的毒性较小。以SKOV3和A549为肿瘤细胞模型，研究了细胞对胶束的摄取。结论叶酸受体高表达的SKOV3细胞对FA-CS-SA的摄取速度明显快于A549细胞，且作用时间短。通过荧光素酶活性检测和绿色荧光蛋白检测，证实FA-CS-SA是一种有效的基因载体。FA-CS-SA/PDNA复合体在SKOV3细胞中的转染率是CS-SA/PDNA复合体的2.3倍。然而，两种复合物在A549细胞中的转染率没有显著差异。重要的是，FA-CS-SA/PDNA在SKOV3细胞中的转染率随游离FA的预处理而降低。结论：FA-CS-SA/PDNA复合体转染率的提高归因于叶酸受体介导的内吞作用。
Background Cationic polymers have been accepted as effective nonviral vectors for gene delivery with low immunogenicity unlike viral vectors. However, the lack of organ or cell specificity sometimes hampers their application and the modification of polymeric vectors has also shown successful improvements in achieving cell-specific targeting delivery and in promoting intracellular gene transfer efficiency. Methods A folic acid-conjugated stearic acid-grafted chitosan (FA-CS-SA) micelle, synthesized by a 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide-coupling reaction, was designed for specific receptor-mediated gene delivery. Results Due to the cationic properties of chitosan, the micelles could compact the plasmid DNA (pDNA) to form micelle/pDNA complexes nanoparticles. The particle size and zeta potential of the FA-CS-SA/pDNA complexes with different N/P ratios were 100–200 nm and −20 to −10 mV, respectively. The DNase I protection assay indicated that the complexes can efficiently protect condensed DNA from enzymatic degradation by DNase I. A cytotoxicity study indicated that the micelles exhibited less toxicity in comparison with LipofectamineTM 2000. Using SKOV3 and A549 as model tumor cells, the cellular uptake of micelles was investigated. Conclusion It was found that cellular uptake of FA-CS-SA in SKOV3 cells with higher folate receptor expression was faster than that in A549 cells with a short incubation time. Luciferase assay and green fluorescent protein detection were used to confirm that FA-CS-SA could be an effective gene vector. Transfection efficiency of the FA-CS-SA/pDNA complexes in SKOV3 cells was enhanced up to 2.3-fold compared with that of the CS-SA/pDNA complexes. However, there was no significant difference between the transfection efficiencies of the two complexes in A549 cells. Importantly, the transfection efficiency of FA-CS-SA/pDNA decreased with free FA pretreatment in SKOV3 cells. It was concluded that the increase in transfection efficiency of the FA-CS-SA/pDNA complexes was attributed to folate receptor-mediated endocytosis.