A biomimetic nanovector-mediated targeted cholesterol-conjugated siRNA delivery for tumor gene therapy
A biomimetic nanovector-mediated targeted cholesterol-conjugated siRNA delivery for tumor gene therapy
复制标题
仿生纳米载体介导的靶向胆固醇缀合 siRNA 递送用于肿瘤基因治疗。
DOI：
10.1016/j.biomaterials.2012.08.057
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发表时间：
2012-12-01
期刊：
影响因子：
14
通讯作者：
Zhang, Qiang
中科院分区：
文献类型：
--
作者：
Ding, Yang;Wang, Wei;Zhang, Qiang
RNA干扰作为一种治疗恶性肿瘤的方法具有巨大的潜力。然而，安全和高效的纳米载体在系统传递小干扰RNA(SiRNA)方面极其缺乏。本研究旨在开发一种仿生纳米载体，重组高密度脂蛋白(RHDL)，介导靶向胆固醇结合的siRNA(Chol-siRNA)，用于精灵宝可梦基因沉默治疗。采用薄膜分散法制备了Chol-siRNA负载的rHDL纳米颗粒(rHDL/Chol-siRNA复合体)，并对其性质进行了详细的研究。在最佳体积比(脂质：Chol-siRNA)下，RHDL/Chol-siRNA复合体具有较高的Chol-siRNA载药率(接近99%)、理想的纳米粒子尺寸和良好的血清稳定性。此外，通过Chol-siRNA释放谱分析，rHDL/Chol-siRNA复合体具有缓释特性和储存稳定性。流式细胞仪和激光共聚焦显微镜观察表明，重组高密度脂蛋白介导的羧基荧光素标记的Chol-siRNA(FAM-Chol-siRNA)可通过高密度脂蛋白受体介导的机制高效地摄取并特异性地将其输送到人肝癌细胞系HepG2中。体外细胞毒性、细胞凋亡率和Western-blotting分析显示，Chol-siRNA-Pokemon纳米粒(rHDL/Chol-siRNA-Pokemon复合体)明显抑制细胞生长，降低细胞中Pokemon和Bcl2蛋白的表达。在体内实验中，近红外染料Cy5标记的Chol-siRNA负载的rHDL纳米粒(rHDL/Cy5-Chol-siRNA复合体)在静脉注射后明显积聚在裸鼠肿瘤内。给药与Cy5-Chol-siRNA载脂复合物(Lipos/Cy5-Chol-siRNA复合体)相比较。此外，rHDL/Chol-siRNA-Pokemon复合体在体内对肿瘤生长有明显的抑制作用，Pokemon和Bcl2蛋白的表达显著降低。这些结果表明，rHDL是一种理想的非病毒肿瘤靶向Chol-siRNA转导载体，rHDL介导的Chol-siRNA-Pokemon转导有望成为肝细胞癌基因治疗的新策略。(C)2012爱思唯尔有限公司。保留所有权利。
RNA interference holds tremendous potential as a therapeutic approach of malignant tumors. However, safe and efficient nanovectors are extremely lack for systemic delivery of small interfering RNA (siRNA). The study aimed to develop a biomimetic nanovector, reconstituted high density lipoprotein (rHDL), mediating targeted cholesterol-conjugated siRNA (Chol-siRNA) delivery for Pokemon gene silencing therapy. Chol-siRNA-loaded rHDL nanoparticles (rHDL/Chol-siRNA complexes) were prepared using thin-film dispersion method and their characteristics were investigated in detail. RHDL/Chol-siRNA complexes at the optimal volume ratio (lipid: Chol-siRNA) exhibited high Chol-siRNA-loading efficiency (similar to 99%), desirable nanoparticle size and excellent stability in serum. In addition, by analyzing Chol-siRNA release profile, rHDL/Chol-siRNA complexes displayed sustained-release characteristic and storage stability. Observations from FACS and confocal microscopic analyses revealed that rHDL-mediated carboxyfluorescein tagged Chol-siRNA (FAM-Chol-siRNA) transfection resulted in highly efficient uptake and specific cytoplasmic delivery of FAM-Chol-siRNA into human hepatocellular carcinoma cell line HepG2 via HDL-receptor mediated mechanism. In vitro cytotoxicity, apoptosis and Western-blot analyses revealed significant cellular growth inhibition and decrease of Pokemon and Bcl-2 protein expression in HepG2 cells treated with Chol-siRNA-Pokemon-loaded rHDL nanoparticles (rHDL/Chol-siRNA-Pokemon complexes), respectively. In in vivo studies, the near-infrared (NIR) dye Cy5 labeled Chol-siRNA-loaded rHDL nanoparticles (rHDL/Cy5-Chol-siRNA complexes) obviously accumulated in tumor of nude mice after i.v. administration as compared with Cy5-Chol-siRNA-loaded lipoplexes (Lipos/Cy5-Chol-siRNA complexes). Morover, rHDL/Chol-siRNA-Pokemon complexes demonstrated great tumor growth inhibition and significant decrease of Pokemon and Bcl-2 protein expression in vivo. These results suggested that rHDL should be an ideal non-viral tumor-targeting vector for Chol-siRNA transfer, and rHDL-mediated Chol-siRNA-Pokemon delivery might be a promising new strategy for gene therapy in hepatocellular carcinoma. (C) 2012 Elsevier Ltd. All rights reserved.