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首页> 外文期刊>African Journal of Biotechnology >Synthesis and characterization of folate-poly(ethylene glycol) chitosan graft-polyethylenimine as a non-viral carrier for tumor-targeted gene delivery
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Synthesis and characterization of folate-poly(ethylene glycol) chitosan graft-polyethylenimine as a non-viral carrier for tumor-targeted gene delivery

机译：叶酸-聚乙二醇壳聚糖接枝-聚乙烯亚胺作为非病毒载体用于肿瘤靶向基因传递的合成与表征

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The use of chitosan and chitosan derivatives for gene delivery is limited due to the low transfection efficiency and difficulty in transfecting into a variety of cell types, including some cancer cells overexpressing folate receptor (FRs). In order to solve this problem, folate (FA) and poly(ethylene glycol) (PEG) was conjugated to chitosan-graft-polyethylenimine (CHI-g-PEI) to enhance water-solubility and the transfection efficiency. In the present study, a cell specific targeting molecule FA was linked on PEG and then grafted the FA-PEG onto CHI-g-PEI. The FA-PEG-grafted CHI-g-PEI (FA-PEG-CHI-g-PEI) effectively condensed the plasmid DNA (pDNA) into nanoparticles with positive surface charge under the suitable nitrogen/phosphorus (N/P) ratio.?In vitro, transfection efficiency of the FA-PEG-CHI-g-PEI /pDNA complex in 293T cells and LoVo cells (FRs over-expressing cell lines) increased with increasing N/P ratio under N/P = 15 and was more than 50%, but no significant difference in human lung carcinoma cells (A549) cells (FRs deficient cell lines). Importantly,?in vivo?luciferase expression showed that the efficiency of FA-PEG-CHI-g-PEI -mediated transfection (50 μg luciferase plasmid (pLuc), N/P ratio = 15) was comparable to that of adenovirus-mediated luciferase transduction (1 × 109pfu) in melanoma-bearing mice. It was concluded that FA-PEG-CHI-g-PEI, which has improved transfection efficiency and FRs specificity?in vitro?and?in vivo, may be useful in gene therapy.
机译：壳聚糖和壳聚糖衍生物用于基因递送的用途受到限制，这是因为转染效率低并且难以转染成多种细胞类型，包括一些过度表达叶酸受体（FRs）的癌细胞。为了解决该问题，将叶酸（FA）和聚（乙二醇）（PEG）与壳聚糖接枝的聚乙烯亚胺（CHI-g-PEI）缀合，以提高水溶性和转染效率。在本研究中，将细胞特异性靶向分子FA连接到PEG上，然后将FA-PEG嫁接到CHI-g-PEI上。在适当的氮/磷（N / P）比下，FA-PEG接枝的CHI-g-PEI（FA-PEG-CHI-g-PEI）有效地将质粒DNA（pDNA）浓缩成具有正表面电荷的纳米颗粒。在体外，FA / PEG-CHI-g-PEI / pDNA复合体在293T细胞和LoVo细胞（FRs过表达的细胞系）中的转染效率随着N / P = 15的N / P比的增加而增加，并且高于50％，但在人肺癌细胞（A549）细胞（FRs缺乏细胞系）中无显着差异。重要的是，体内荧光素酶表达表明，FA-PEG-CHI-g-PEI介导的转染效率（50μg荧光素酶质粒（pLuc），N / P比= 15）与腺病毒介导的荧光素酶相当荷黑素瘤小鼠的转导（1×109pfu）。结论是，FA-PEG-CHI-g-PEI具有更高的体外和体内转染效率和FRs特异性，可用于基因治疗。

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