Protein Research Group and §MEMPHYS Center for Biomembrane Physics, Department of Biochemistry and Molecular Biology, University of Southern Denmark , Odense, DK-5230, Denmark.
ACS Nano. 2014 Mar 25;8(3):2161-75. doi: 10.1021/nn4050744. Epub 2014 Feb 20.
The use of nanoparticles in foods, materials, and clinical treatments has increased dramatically in the past decade. Because of the possibility of human exposure to nanoparticles, there is an urgent need to investigate the molecular mechanisms underlying the cellular responses that might be triggered. Such information is necessary to assess potential health risks arising from the use of nanoparticles, and for developing new formulations of next generation nanoparticles for clinical treatments. Using mass spectrometry-based proteomic technologies and complementary techniques (e.g., Western blotting and confocal laser scanning microscopy), we present insights into the silver nanoparticle-protein interaction in the human LoVo cell line. Our data indicate that some unique cellular processes are driven by the size. The 100 nm nanoparticles exerted indirect effects via serine/threonine protein kinase (PAK), mitogen-activated protein kinase (MAPK), and phosphatase 2A pathways, and the 20 nm nanoparticles induced direct effects on cellular stress, including generation of reactive oxygen species and protein carbonylation. In addition, we report that proteins involved in SUMOylation were up-regulated after exposure to 20 nm silver nanoparticles. These results were further substantiated by the observation of silver nanoparticles entering the cells; however, data indicate that this was determined by the size of the nanoparticles, since 20 nm particles entered the cells while 100 nm particles did not.
在过去十年中，纳米粒子在食品、材料和临床治疗中的应用有了显著的增长。由于人类可能接触到纳米粒子，因此迫切需要研究可能引发细胞反应的分子机制。这些信息对于评估纳米粒子的使用可能带来的健康风险，以及开发用于临床治疗的下一代纳米粒子的新配方是必要的。本研究采用基于质谱的蛋白质组学技术和互补技术（如 Western blot 和共聚焦激光扫描显微镜），深入研究了人 LoVo 细胞系中银纳米粒子与蛋白质的相互作用。我们的数据表明，一些独特的细胞过程是由粒径驱动的。100nm 纳米粒子通过丝氨酸/苏氨酸蛋白激酶（PAK）、丝裂原活化蛋白激酶（MAPK）和磷酸酶 2A 途径发挥间接作用，而 20nm 纳米粒子则对细胞应激直接产生影响，包括活性氧的产生和蛋白质羰基化。此外，我们还报告称，暴露于 20nm 银纳米粒子后，与 SUMO 化相关的蛋白质被上调。这些结果进一步通过观察到银纳米粒子进入细胞得到证实；然而，数据表明，这是由纳米粒子的粒径决定的，因为 20nm 颗粒进入了细胞，而 100nm 颗粒则没有。
