title: Protein transfection study using multicellular tumor spheroids of human hepatoma Huh-7 cells.
abstract: 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.
authors.affiliation: Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
authors.forename: Takuma
authors.initials: T
authors.lastname: Kato
authors.affiliation: Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Tanaka
authors.affiliation: Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Oba
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000473:pathology / Q000628:therapy; D045744:Cell Line, Tumor; D016503:Drug Delivery Systems; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000628:therapy; D018874:Spheroids, Cellular / Q000378:metabolism* / Q000473:pathology