He Bo, Yang Dan, Qin Mengmeng, Zhang Yuan, He Bing, Dai Wenbing, Wang Xueqing, Zhang Qiang, Zhang Hua, Yin Changcheng
Department of Biophysics, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery Systems, School of Pharmaceutical Sciences, Peking University, Beijing, China; State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, China.
Biochem Biophys Res Commun. 2017 Dec 9;494(1-2):339-345. doi: 10.1016/j.bbrc.2017.10.026. Epub 2017 Oct 7.
Gold nanoparticles are promising drug delivery vehicles for nucleic acids, small molecules, and proteins, allowing various modifications on the particle surface. However, the instability and low bioavailability of gold nanoparticles compromise their clinical application. Here, we functionalized gold nanoparticles with CPP fragments (CALNNPFVYLI, CALRRRRRRRR) through sulfhydryl PEG to increase their stability and bioavailability. The resulting gold nanoparticles were characterized with transmission electron microscopy (TEM), dynamic light scattering (DLS), UV-visible spectrometry and X-ray photoelectron spectroscopy (XPS), and the stability in biological solutions was evaluated. Comparing to PEGylated gold nanoparticles, CPP (CALNNPFVYLI, CALRRRRRRRR)-modified gold nanoparticles showed 46 folds increase in cellular uptake in A549 and B16 cell lines, as evidenced by the inductively coupled plasma atomic emission spectroscopy (ICP-AES). The interactions between gold nanoparticles and liposomes indicated CPP-modified gold nanoparticles bind to cell membrane more effectively than PEGylated gold nanoparticles. Surface plasmon resonance (SPR) was used to measure interactions between nanoparticles and the membrane. TEM and uptake inhibitor experiments indicated that the cellular entry of gold nanoparticles was mediated by clathrin and macropinocytosis. Other energy independent endocytosis pathways were also identified. Our work revealed a new strategy to modify gold nanoparticles with CPP and illustrated the cellular uptake pathway of CPP-modified gold nanoparticles.
金纳米颗粒是用于核酸、小分子和蛋白质的很有前景的药物递送载体，可在颗粒表面进行各种修饰。然而，金纳米颗粒的不稳定性和低生物利用度限制了它们的临床应用。在这里，我们通过巯基聚乙二醇用细胞穿透肽片段（CALNNPFVYLI、CALRRRRRRRR）对金纳米颗粒进行功能化，以提高其稳定性和生物利用度。通过透射电子显微镜（TEM）、动态光散射（DLS）、紫外可见光谱和X射线光电子能谱（XPS）对所得金纳米颗粒进行了表征，并评估了其在生物溶液中的稳定性。与聚乙二醇化金纳米颗粒相比，细胞穿透肽（CALNNPFVYLI、CALRRRRRRRR）修饰的金纳米颗粒在A549和B16细胞系中的细胞摄取增加了46倍，这通过电感耦合等离子体原子发射光谱（ICP-AES）得到证实。金纳米颗粒与脂质体之间的相互作用表明，细胞穿透肽修饰的金纳米颗粒比聚乙二醇化金纳米颗粒更有效地结合到细胞膜上。表面等离子体共振（SPR）用于测量纳米颗粒与膜之间的相互作用。透射电子显微镜和摄取抑制剂实验表明，金纳米颗粒的细胞内吞是由网格蛋白和巨胞饮作用介导的。还确定了其他不依赖能量的内吞途径。我们的工作揭示了一种用细胞穿透肽修饰金纳米颗粒的新策略，并阐明了细胞穿透肽修饰的金纳米颗粒的细胞摄取途径。
