Redox-Sensitive PEG-Polypeptide Nanoporous Particles for Survivin Silencing in Prostate Cancer Cells
Redox-Sensitive PEG-Polypeptide Nanoporous Particles for Survivin Silencing in Prostate Cancer Cells
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DOI：
10.1021/acs.biomac.5b00562
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发表时间：
2015-07-01
影响因子：
6.2
通讯作者：
Caruso, Frank
中科院分区：
文献类型：
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作者：
Cavalieri, Francesca;Beretta, Giovanni L.;Caruso, Frank
我们报告了细胞内氧化还原响应性纳米多孔聚（乙二醇）聚（L-赖氨酸）颗粒（NPEG-PLL）的工程。所获得的颗粒表现出无毒性，同时保持递送靶向前列腺癌细胞中的抗凋亡因子生存素的小干扰RNA序列（siRNA）的能力。氧化还原介导的稳定NPEG-PLL-siRNA复合物的二硫键裂解导致生物活性siRNA释放到前列腺癌PC-3细胞的胞质溶胶中，这反过来又导致靶基因的有效沉默（类似于59 +/-8%）。在最佳条件下获得的这些发现表明，NPEG-PLL可以保护细胞外和细胞内环境中的治疗性核酸，从而防止与血清和胞质蛋白的竞争性相互作用的发生以及RNA酶的降解。通过去卷积显微镜、荧光寿命成像显微镜和超分辨率结构照明显微镜的组合研究NPEG-PLL的细胞内运输和最终命运。在同时暴露于紫杉醇和siRNA负载的NPEG-PLL的细胞中观察到细胞存活的显著损害。总的来说，我们的研究结果表明，NPEG-PLL代表了用于将治疗性核酸递送至癌细胞的高负载贮库。
We report the engineering of intracellular redox-responsive nanoporous poly(ethylene glycol) poly(L-lysine) particles (NPEG-PLLs). The obtained particles exhibit no toxicity while maintaining the capability to deliver a small interfering RNA sequence (siRNA) targeting the anti-apoptotic factor, survivin, in prostate cancer cells. The redox-mediated cleavage of the disulfide bonds stabilizing the NPEG-PLL-siRNA complex results in the release of bioactive siRNA into the cytosol of prostate cancer PC-3 cells, which, in turn, leads to the effective silencing (similar to 59 +/- 8%) of the target gene. These findings, obtained under optimal conditions, indicate that NPEG-PLLs may protect the therapeutic nucleic acid in the extracellular and intracellular environments, thus preventing the occurrence of competitive interactions with serum and cytosolic proteins as well as degradation by RNase. The intracellular trafficking and final fate of the NPEG-PLLs were investigated by a combination of deconvolution microscopy, fluorescence lifetime imaging microscopy, and super-resolution structured illumination microscopy. A significant impairment of cell survival was observed in cells concomitantly exposed to paclitaxel and siRNA-loaded NPEG-PLLs. Overall, our findings indicate that NPEG-PLLs represent a highly loaded depot for the delivery of therapeutic nucleic acids to cancer cells.