Delivery system for DNAzymes using arginine-modified hydroxyapatite nanoparticles for therapeutic application in a nasopharyngeal carcinoma model.
Delivery system for DNAzymes using arginine-modified hydroxyapatite nanoparticles for therapeutic application in a nasopharyngeal carcinoma model.
复制标题
使用精氨酸修饰的羟基磷灰石纳米粒子的 DNAzyme 递送系统在鼻咽癌模型中的治疗应用
DOI：
10.2147/ijn.s48321
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发表时间：
2013
影响因子：
8
通讯作者：
Sun L
中科院分区：
文献类型：
--
作者：
Chen Y;Yang L;Huang S;Li Z;Zhang L;He J;Xu Z;Liu L;Cao Y;Sun L
DNAzyme是一种人工合成的单链催化核酸，它以序列特异性的方式结合和切割目标mRNA，并已被探索用于基因治疗。制约它们应用的一个瓶颈是缺乏有效的交付系统。纳米羟基磷灰石(NHAP)作为一种具有潜在应用前景的无机纳米材料，作为非病毒载体的新候选材料，受到了越来越多的关注。在这项研究中，我们开发了一种基于nHAP的递送系统，并探索了它的细胞摄取机制、细胞内定位和生物学效应。体外条件下，精氨酸修饰的纳米羟基磷灰石颗粒(Arg-nHAP)和DZ1(潜伏膜蛋白1[LMP1]靶向)的吸附效率接近100%。使用特定的抑制剂，细胞摄取Arg-nHAP/DZ1复合体被证明是通过能量依赖的内吞途径介导的。此外，通过共聚焦显微镜证实了该复合体有效的细胞内递送和核定位。生物学上，该复合体成功地下调了鼻咽癌细胞中LMP1的表达。在小鼠肿瘤异种移植模型中，该复合体被证明能有效地输送到肿瘤组织，下调LMP1的表达，抑制肿瘤生长。这些结果表明，Arg-nHAP可能是一种具有潜在临床应用前景的核酸药物载体。
DNAzymes are synthetic, single-stranded, catalytic nucleic acids that bind and cleave target mRNA in a sequence-specific manner, and have been explored for genotherapeutics. One bottleneck restricting their application is the lack of an efficient delivery system. As an inorganic nanomaterial with potentially wide application, nano-hydroxyapatite particles (nHAP) have attracted increasing attention as new candidates for nonviral vectors. In this study, we developed an nHAP-based delivery system and explored its cellular uptake mechanisms, intracellular localization, and biological effects. Absorption of arginine-modified nanohydroxyapatite particles (Arg-nHAP) and DZ1 (latent membrane protein 1 [LMP1]-targeted) reached nearly 100% efficiency under in vitro conditions. Using specific inhibitors, cellular uptake of the Arg-nHAP/DZ1 complex was shown to be mediated by the energy-dependent endocytosis pathway. Further, effective intracellular delivery and nuclear localization of the complex was confirmed by confocal microscopy. Biologically, the complex successfully downregulated the expression of LMP1 in nasopharyngeal carcinoma cells. In a mouse tumor xenograft model, the complex was shown to be delivered efficiently to tumor tissue, downregulating expression of LMP1 and suppressing tumor growth. These results suggest that Arg-nHAP may be an efficient vector for nucleic acid-based drugs with potential clinical application.