Intracellular delivery and anti-cancer effect of self-assembled heparin-Pluronic nanogels with RNase A
Intracellular delivery and anti-cancer effect of self-assembled heparin-Pluronic nanogels with RNase A
复制标题
DOI：
10.1016/j.jconrel.2010.07.118
复制
发表时间：
2010-11-01
影响因子：
10.8
通讯作者：
Park, Ki Dong
中科院分区：
文献类型：
--
作者：
Choi, Jong Hoon;Jang, Ji Young;Park, Ki Dong
制备了一种新型的自组装纳米凝胶，用于核糖核酸酶A(RNase A)的细胞内传递，并研究了RNase A的抗癌效果。用动态光散射(DLS)、Xi-电位和透射电子显微镜(TEM)表征了含RNaseA的肝素-Pluronic(HP)自组装纳米凝胶(HPR纳米凝胶)的物理性质。核糖核酸酶A对HP纳米凝胶有很强的亲和力，具有很高的负载效率(>；78%)，并显著减小了流体力学尺寸(从89降至-29)。HPR纳米凝胶能有效地内化到HeLa细胞中，并定位于胞浆和胞核。在细胞摄取机制的研究中，甲氧基-β-环糊精(M-β-CD)降低了HP纳米凝胶的摄取效率，表明内化是通过小窝/脂筏介导的内吞作用发生的。核内的定位很可能是因为结合肝素促进了核的穿透。随着RNaseA浓度的增加，HPR纳米凝胶的细胞毒性显著增强，这是由于HPR纳米凝胶的细胞内定位导致胞浆和细胞核中单链RNA的降解所致。这些结果表明，自组装的HP纳米凝胶是一种非凡的细胞内蛋白输送工具，有望用作癌症化疗药物。(C)2010爱思唯尔B.V.保留所有权利。
A novel self-assembled nanogel was prepared for the intracellular delivery of ribonuclease A (RNase A) and the anti-cancer efficacy of RNase A delivery was investigated. The physical properties of self-assembled heparin-Pluronic (HP) nanogels incorporating RNase A (HPR nanogels) were characterized by dynamic light scattering (DLS), xi-potential, and transmission electron microscopy (TEM). RNase A showed a strong affinity for the HP nanogel, resulting in a high loading efficiency (> 78%) and significantly decreased hydrodynamic size (from 89 to -29). HPR nanogels were efficiently internalized into HeLa cells and localized in the cytosol as well as the nucleus. In the mechanism study of cellular uptake, treating with methoxy beta-cyclodextrin (M beta-CD) decreased the uptake efficiency of HP nanogel, indicating that internalization occurs via caveolae/lipid-raft mediated endocytosis. Localization in the nucleus most likely occurred because the conjugated heparin facilitated nucleus penetration. The cytotoxicity of HPR nanogels was significantly increased when the RNase A concentration was increased, which resulted from the degradation of single stranded RNAs in the cytosol and the nucleus due to the intracellular localization of the HPR nanogels. These results demonstrate that self-assembled HP nanogels are a remarkable vehicle for intracellular protein delivery and hold promise for use as cancer chemotherapeutics. (C) 2010 Elsevier B.V. All rights reserved.