Tekle Christina, Deurs Bo van, Sandvig Kirsten, Iversen Tore-Geir
Centre for Cancer Biomedicine, Faculty Division, Norwegian Radium Hospital, University of Oslo, 0316 Oslo, Norway.
Nano Lett. 2008 Jul;8(7):1858-65. doi: 10.1021/nl0803848. Epub 2008 Jun 21.
Can quantum dots (Qdots) act as relevant intracellular probes to investigate routing of ligands in live cells? The intracellular trafficking of Qdots that were coupled to the plant toxin ricin, Shiga toxin, or the ligand transferrin (Tf) was studied by confocal fluorescence microscopy. The Tf:Qdots were internalized by clathrin-dependent endocytosis as fast as Tf, but their recycling was blocked. Unlike Shiga toxin, the Shiga:Qdot bioconjugate was not routed to the Golgi apparatus. The internalized ricin:Qdot bioconjugates localized to the same endosomes as ricin itself but could not be visualized in the Golgi apparatus. Importantly, we find that the endosomal accumulation of ricin:Qdots affects endosome-to-Golgi transport of both ricin and Shiga toxin: Transport of ricin was reduced whereas transport of Shiga toxin was increased. In conclusion, the data reveal that, although coupling of Qdots to a ligand does not necessarily change the endocytic pathway normally used by the ligands studied, it appears that the ligand-coupled Qdot nanoparticles can be arrested within endosomes and somehow perturb the normal endosomal sorting in cells. Thus, the results demonstrate that Qdots may have severe consequences on cell physiology.
量子点(Qdots)能否作为相关的细胞内探针来研究活细胞中配体的转运途径?通过共聚焦荧光显微镜研究了与植物毒素蓖麻毒素、志贺毒素或配体转铁蛋白(Tf)偶联的量子点在细胞内的运输情况。转铁蛋白:量子点(Tf:Qdots)通过网格蛋白依赖的内吞作用内化的速度与转铁蛋白一样快,但它们的再循环被阻断。与志贺毒素不同,志贺毒素:量子点生物共轭物不会被转运到高尔基体。内化的蓖麻毒素:量子点生物共轭物定位于与蓖麻毒素本身相同的内体,但在高尔基体中无法观察到。重要的是,我们发现蓖麻毒素:量子点在内体中的积累会影响蓖麻毒素和志贺毒素从内体到高尔基体的运输:蓖麻毒素的运输减少,而志贺毒素的运输增加。总之,数据表明,虽然量子点与配体的偶联不一定会改变所研究配体通常使用的内吞途径,但似乎配体偶联的量子点纳米颗粒可以在内体中滞留,并以某种方式扰乱细胞内正常的内体分选。因此,结果表明量子点可能对细胞生理学产生严重影响。