Tat peptide mediated cellular uptake of SiO2 submicron particles.
Tat peptide mediated cellular uptake of SiO2 submicron particles.
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DOI：
10.1016/j.colsurfb.2009.09.017
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发表时间：
2010-02
期刊：
影响因子：
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通讯作者：
Z. Mao;L. Wan;Ling Hu;Lie Ma;Changyou Gao
中科院分区：
文献类型：
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作者：
Z. Mao;L. Wan;Ling Hu;Lie Ma;Changyou Gao
纳米粒子和微粒子在活细胞中的内在化与其潜在的应用、功能、细胞毒性和细胞内药物传递密切相关。特别是，蛋白质、多肽、核酸和小颗粒等大量货物进入细胞可以通过一些配体（如Tat肽）来增强。在这项工作中，评估了Tat介导的细胞摄取能力。将Tat肽共价固定在直径为200nm的荧光素标记的sio2颗粒（fitc - sio2 - nh2颗粒）上。BCA蛋白测定表明，Tat的投料量可控制在0 ~ 3.5μg/mg sio2颗粒的接枝量范围内。Tat肽的表面固定对sio2 - nh2颗粒的表面形貌和电荷性质没有明显的影响。相反，fitc - sio2 - nh2颗粒和FITC-SiO2-Tat颗粒的表面电荷在Dulbecco's Modified Eagles Medium （DMEM）中呈微阳性，在DMEM/胎牛血清中呈微阴性，传递了血浆蛋白在颗粒上的吸附。流式细胞术检测显示，FITC-SiO2-Tat颗粒被HepG2细胞内化的速度明显快于fitc - sio2 - nh2颗粒，颗粒数量也明显多于fitc - sio2 - nh2颗粒。此外，4℃低温对Tat肽修饰颗粒内化的影响较小。Tat修饰也影响了颗粒的亚细胞分布，导致颗粒在细胞核内的定位。fitc - sio2 - nh2颗粒和FITC-SiO2-Tat颗粒均未检测到明显的细胞毒性。
Internalization of nano- and microparticles into live cells correlates closely with their potential applications, functions, cytotoxicity and intracellular drug delivery. Particularly, delivery of a large variety of cargoes such as proteins, peptides, nucleic acids and small particles into cells could be enhanced by some ligands such as Tat peptide. In this work, the ability of Tat mediated cellular uptake was assessed. The Tat peptide was covalently immobilized to fluorescein tagged SiO2particles (FITC–SiO2–NH2particles) with a diameter of 200nm. BCA protein assay determined that the grafting amount of the Tat peptide could be controlled within a range of 0–3.5μg/mg SiO2particles by the Tat feeding amount. Surface immobilization of the Tat peptide did not bring apparent changes on the surface morphology and charge property of the SiO2–NH2particles. By contrast, the surface charge of both the FITC–SiO2–NH2particles and the FITC–SiO2–Tat particles was reversed from slight positive in Dulbecco's Modified Eagles Medium (DMEM) to slight negative in DMEM/fetal bovine serum, conveying adsorption of plasma proteins on the particles. Flow cytometry measurement showed that the FITC–SiO2–Tat particles were internalized by HepG2 cells with a significant faster rate and a higher number of particles than that of the FITC–SiO2–NH2particles. Moreover, internalization of the Tat peptide decorated particles was less influenced by the low temperature at 4°C. The Tat decoration affected the subcellular distribution of the particles as well, resulting in localization of the particles in the cell nucleus. No obvious cytotoxicity was detected for both the FITC–SiO2–NH2particles and the FITC–SiO2–Tat particles.