State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University , 24 Tongjiaxiang, Nanjing 210009, China.
School of Pharmaceutical Science, Nanjing Tech University , 30 South Puzhu Road, Nanjing 211816, China.
ACS Appl Mater Interfaces. 2017 Jul 19;9(28):23353-23369. doi: 10.1021/acsami.7b04971. Epub 2017 Jul 5.
Single-walled carbon nanotubes (SWNTs) with unique physicochemical properties have exhibited promising biomedical applications as drug and gene carriers. In this study, polyethylenimine (PEI)-modified SWNT conjugates linked with candesartan (CD) were developed to deliver vascular endothelial growth factor (VEGF)-targeted siRNA (siVEGF) for the synergistic and targeted treatment of tumor angiogenesis. The characterization results revealed that SWNT-PEI-CD conjugates were successfully synthesized and exhibited desirable dispersibility and superior stability. Confocal laser scanning microscopy (CLSM) and flow cytometry (FCM) results showed that SWNT-PEI-CD/siVEGF complexes could achieve high cellular uptake and specific intracellular distribution of siRNA in ATR overexpressed PANC-1 cells. Strong down-regulation of VEGF was also verified by qualitative real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blot in complex-treated PANC-1 cells. The in vitro angiogenesis assay showed that SWNT-PEI-CD/siVEGF complexes highly inhibited tube formation of human umbilical vein endothelial cells. Furthermore, in vivo observation in PANC-1 xenografted nude mice demonstrated that SWNT-PEI-CD/siVEGF complexes exhibited significant distribution at tumor sites and caused obvious inhibition of tumor growth and tumor-associated angiogenesis repression induced by the drug combination of CD and siVEGF. Finally, a WST-1 assay indicated that SWNT-PEI-CD possessed low cytotoxicity, and a hemolysis test showed good biocompatibility of SWNT-PEI-CD. Hematological and histological analyses confirmed that SWNT-PEI-CD/siVEGF complexes did not cause any obvious toxic effects to blood and major organs. These findings suggested that the SWNT-PEI-CD/siVEGF co-delivery system with tumor-targeting specificity, improved endosomal escaping properties, and collaboration of angiogenesis inhibition could be a prospective method for efficient tumor antiangiogenic therapy.
单壁碳纳米管（SWNTs）具有独特的物理化学性质，已作为药物和基因载体在生物医学领域展现出广阔的应用前景。本研究制备了聚乙二烯亚胺（PEI）修饰的SWNT 与坎地沙坦（CD）偶联物，用于递送血管内皮生长因子（VEGF）靶向 siRNA（siVEGF），以协同靶向治疗肿瘤血管生成。表征结果表明，成功合成了 SWNT-PEI-CD 偶联物，其具有良好的分散性和优异的稳定性。共聚焦激光扫描显微镜（CLSM）和流式细胞术（FCM）结果表明，SWNT-PEI-CD/siVEGF 复合物能够实现 ATR 过表达的 PANC-1 细胞内 siRNA 的高细胞摄取和特异性细胞内分布。复合物处理的 PANC-1 细胞中，通过定性实时聚合酶链反应、酶联免疫吸附试验和 Western blot 也验证了 VEGF 的强烈下调。体外血管生成实验表明，SWNT-PEI-CD/siVEGF 复合物高度抑制人脐静脉内皮细胞的管形成。此外，在 PANC-1 异种移植裸鼠体内观察到，SWNT-PEI-CD/siVEGF 复合物在肿瘤部位有明显的分布，并导致由 CD 和 siVEGF 联合用药引起的肿瘤生长和肿瘤相关血管生成抑制明显抑制。最后，WST-1 测定表明 SWNT-PEI-CD 具有低细胞毒性，溶血试验表明 SWNT-PEI-CD 具有良好的生物相容性。血液学和组织学分析证实，SWNT-PEI-CD/siVEGF 复合物对血液和主要器官没有造成明显的毒性作用。这些发现表明，具有肿瘤靶向特异性、提高内体逃逸性能和协同抗血管生成作用的 SWNT-PEI-CD/siVEGF 共递药系统可能成为高效肿瘤抗血管生成治疗的有前途的方法。
