Kato Takuma, Tanaka Masakazu, Oba Makoto
Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
PLoS One. 2013 Dec 5;8(12):e82876. doi: 10.1371/journal.pone.0082876. eCollection 2013.
Several protein transfection reagents are commercially available and are powerful tools for elucidating function of a protein in a cell. Here we described protein transfection studies of the commercially available reagents, Pro-DeliverIN, Xfect, and TuboFect, using Huh-7 multicellular tumor spheroid (MCTS) as a three-dimensional in vitro tumor model. A cellular uptake study using specific endocytosis inhibitors revealed that each reagent was internalized into Huh-7 MCTS by different mechanisms, which were the same as monolayer cultured Huh-7 cells. A certain amount of Pro-DeliverIN and Xfect was uptaken by Huh-7 cells through caveolae-mediated endocytosis, which may lead to transcytosis through the surface-first layered cells of MCTS. The results presented here will help in the choice and use of protein transfection reagents for evaluating anti-tumor therapeutic proteins against MCTS models.
有几种蛋白质转染试剂在市场上有售，它们是阐明蛋白质在细胞中功能的有力工具。在此，我们描述了使用Pro-DeliverIN、Xfect和TuboFect这几种市售试剂，以Huh-7多细胞肿瘤球体（MCTS）作为三维体外肿瘤模型进行的蛋白质转染研究。一项使用特异性内吞抑制剂的细胞摄取研究表明，每种试剂通过不同机制被内化到Huh-7 MCTS中，这与单层培养的Huh-7细胞相同。一定量的Pro-DeliverIN和Xfect通过小窝介导的内吞作用被Huh-7细胞摄取，这可能导致通过MCTS表面第一层细胞的转胞吞作用。本文给出的结果将有助于选择和使用蛋白质转染试剂来评估针对MCTS模型的抗肿瘤治疗性蛋白质。
