Significance of the balance between intracellular glutathione and polyethylene glycol for successful release of small interfering RNA from gold nanoparticles
Significance of the balance between intracellular glutathione and polyethylene glycol for successful release of small interfering RNA from gold nanoparticles
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DOI：
10.1007/s12274-015-0828-5
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发表时间：
2015-10-01
期刊：
影响因子：
9.9
通讯作者：
Berry, Catherine C.
中科院分区：
文献类型：
--
作者：
McCully, Mark;Hernandez, Yulan;Berry, Catherine C.
小干扰rna （sirna）用于特定基因沉默的治疗前景依赖于功能性sirna成功递送到细胞质。它们结合到一个既定的传递平台上，如金纳米颗粒，为治疗疾病和促进我们对细胞过程的理解提供了巨大的潜力。它们的成功或失败取决于纳米颗粒进入细胞的摄取和随后细胞内功能性siRNA的释放。在这项研究中，我们利用金纳米颗粒sirna介导的C-MYC递送，旨在确定我们是否可以在细胞内谷胱甘肽水平较低的癌细胞系中实现敲除，并确定聚乙二醇（PEG）配体密度对敲除的影响，以确定实现C-MYC敲除的最佳纳米颗粒设计。我们证明，无论PEG密度如何，在相对较低谷胱甘肽水平的细胞中都可以实现敲低，以及PEG的空间位阻对细胞内环境中siRNA切割可用性的可能影响。通过透射电子显微镜和质谱分析证明了金纳米颗粒的摄取，而通过细胞内western和BrdU掺入分别在蛋白质和生理水平（s期细胞）上确定了敲低。
The therapeutic promise of small interfering RNAs (siRNAs) for specific gene silencing is dependent on the successful delivery of functional siRNAs to the cytoplasm. Their conjugation to an established delivery platform, such as gold nanoparticles, offers tremendous potential for treating diseases and advancing our understanding of cellular processes. Their success or failure is dependent on both the uptake of the nanoparticles into the cells and subsequent intracellular release of the functional siRNA. In this study, utilizing gold nanoparticle siRNA-mediated delivery against C-MYC, we aimed to determine if we could achieve knockdown in a cancer cell line with low levels of intracellular glutathione, and determine the influence, if any, of polyethylene glycol (PEG) ligand density on knockdown, with a view to determining the optimal nanoparticle design to achieve C-MYC knockdown. We demonstrate that, regardless of the PEG density, knockdown in cells with relatively low glutathione levels can be achieved, as well as the possible effect of steric hindrance of PEG on the availability of the siRNA for cleavage in the intracellular environment. Gold nanoparticle uptake was demonstrated via transmission electron microscopy and mass spectroscopy, while knockdown was determined at the protein and physiological levels (cells in S-phase) by in-cell westerns and BrdU incorporation, respectively.