Key Laboratory of Biomedical Polymers of Ministry of Education and Department of Chemistry, Wuhan University , Wuhan 430072, People's Republic of China.
ACS Appl Mater Interfaces. 2017 Dec 13;9(49):42622-42632. doi: 10.1021/acsami.7b14963. Epub 2017 Dec 1.
A facile and targeted gene delivery system was prepared by conjugating β-cyclodextrin modified polyethylenimine (PEI-CD) and adamantyl peptide (AdGRGDS) based on host-guest interaction. With the rational design between PEI-CD and AdGRGDS, the PEI-CD/AdGRGDS gene delivery system showed excellent DNA binding capability and exhibited good ability to compact DNA into uniform spherical nanoparticles. In vitro luciferase assay showed that gene expression transfected by PEI-CD/AdGRGDS was stronger than that by PEI-CD in HeLa cells, whereas gene expression transfected by PEI-CD/AdGRGDS and PEI-CD was similar to each other in COS7 cells. Internalization of complexes was qualitatively studied using a confocal laser scanning microscope (CLSM) and quantitatively analyzed by flow cytometry, respectively, and targeting specificity was also evaluated by CLSM. Results of CLSM and flow cytometry indicated that PEI-CD/AdGRGDS had good targeting specificity to tumor cells with integrin αvβ3 overexpression. To further evaluate the targeting specificity and transfection efficiency in vivo, a rat model with murine hepatic carcinoma cell line H22 was used. PEI-CD/AdGRGDS showed stronger gene expression efficiency than PEI-CD via in vivo transfection of pORF-LacZ and pGL-3 plasmids after subcutaneous injection. Interestingly, PEI-CD/AdGRGDS also showed high targeting specificity and transfection distribution to tumor xenograft after tail-vein injection. In vitro and in vivo assays highlighted the importance of GRGDS targeting specificity to tumor cells with integrin αvβ3 overexpression and demonstrated that the PEI-CD/AdGRGDS gene delivery system would have great potential for targeted tumor therapy.
一种基于主客体相互作用的简便靶向基因传递系统，通过将β-环糊精修饰的聚乙烯亚胺（PEI-CD）与金刚烷肽（AdGRGDS）偶联制备而成。通过合理设计 PEI-CD 和 AdGRGDS，PEI-CD/AdGRGDS 基因传递系统表现出优异的 DNA 结合能力，并能够将 DNA 有效地压缩成均匀的球形纳米颗粒。体外荧光素酶报告基因实验结果表明，PEI-CD/AdGRGDS 转染的基因表达活性在 HeLa 细胞中强于 PEI-CD，而在 COS7 细胞中 PEI-CD/AdGRGDS 和 PEI-CD 的基因表达活性相似。利用共聚焦激光扫描显微镜（CLSM）对复合物的内化进行定性研究，用流式细胞术进行定量分析，并通过 CLSM 评价其靶向特异性。CLSM 和流式细胞术的结果表明，PEI-CD/AdGRGDS 对整合素 αvβ3 过表达的肿瘤细胞具有良好的靶向特异性。为了进一步评价其在体内的靶向特异性和转染效率，构建了荷鼠肝癌细胞系 H22 的大鼠模型。皮下注射 pORF-LacZ 和 pGL-3 质粒后，PEI-CD/AdGRGDS 的体内转染显示出比 PEI-CD 更强的基因表达效率。有趣的是，尾静脉注射后，PEI-CD/AdGRGDS 也显示出对肿瘤异种移植物的高靶向特异性和转染分布。体内外实验结果强调了 GRGDS 靶向整合素 αvβ3 过表达肿瘤细胞的特异性，并表明 PEI-CD/AdGRGDS 基因传递系统在靶向肿瘤治疗方面具有很大的潜力。
