Institute of Nanochemistry and Nanobiology, Shanghai University, Shanghai 200444, China.
Institute of Nanochemistry and Nanobiology, Shanghai University, Shanghai 200444, China; Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
Colloids Surf B Biointerfaces. 2021 Dec;208:112140. doi: 10.1016/j.colsurfb.2021.112140. Epub 2021 Sep 25.
Semiconductor quantum dots (QDs) have been extensively explored for extensive bioapplications, yet their cellular fate, especially exocytosis, has not been thoroughly investigated. Herein, we systematically investigated the whole cellular process from the endocytosis, intercellular trafficking, to the exocytosis of a typical QD, core/shell CdSe/ZnS QD. Using confocal laser scanning microscopy and flow cytometry, and after carefully eliminating the effect of cell division, we found that the QDs were internalized by HeLa cells with a time-, dose-, and serum-dependent manner. The cellular uptake was inhibited by serum, but eventually peaked after 4-6 h incubation with or without serum. The primary endocytosis pathway was clathrin-mediated, and actin- and microtubule-dependent in the medium with serum, while the caveolae-mediated endocytosis and macropinocytosis were more important for the QDs in the serum-free medium. Inside cells, most QDs distributed in lysosomes, and some entered mitochondria, endoplasmic reticulum, and Golgi apparatus. The translocation of the QDs from other organelles to Golgi apparatus was observed. The exocytosis of QDs was faster than the endocytosis, reaching the maximum in about one hour after cultured in fresh culture medium, with around 60% of the internalized QDs remained undischarged. The exocytosis process was energy- and actin-dependent, and the lysosome exocytosis and endoplasmic reticulum/Golgi pathway were the main routes. This study provides a full picture of behavior and fate of QDs in cells, which may facilitate the design of ideal QDs applied in biomedical and other fields.
半导体量子点(QDs)在广泛的生物应用中得到了广泛的研究,但其细胞命运,特别是胞吐作用,尚未得到彻底研究。在此,我们系统地研究了典型 QD(核/壳 CdSe/ZnS QD)从内吞作用、细胞间转运到胞吐作用的整个细胞过程。通过共聚焦激光扫描显微镜和流式细胞术,并仔细消除细胞分裂的影响,我们发现 QD 以时间、剂量和血清依赖的方式被 HeLa 细胞内化。血清抑制细胞摄取,但最终在有或没有血清孵育 4-6 小时后达到峰值。初级内吞作用途径是网格蛋白介导的,在含血清的培养基中依赖于肌动蛋白和微管,而无血清培养基中更重要的是小窝蛋白介导的内吞作用和巨胞饮作用。在细胞内,大多数 QD 分布在溶酶体中,一些进入线粒体、内质网和高尔基体。观察到 QD 从其他细胞器向高尔基体的转运。QD 的胞吐作用比内吞作用快,在新鲜培养基中培养约 1 小时后达到最大值,约 60%的内化 QD 未排出。胞吐作用过程依赖于能量和肌动蛋白,溶酶体胞吐作用和内质网/高尔基体途径是主要途径。这项研究提供了 QD 在细胞内的行为和命运的全貌,这可能有助于设计应用于生物医学和其他领域的理想 QD。