title: Cell line-dependent internalization pathways determine DNA transfection efficiency of decaarginine-PEG-lipid.
abstract: Previously, we have reported that decaarginine-conjugated PEG-lipids (R10B) efficiently delivered plasmid DNA (pDNA) into human cervical carcinoma HeLa cells via macropinocytosis; however, the mechanism of cellular uptake by R10B was not evaluated in other cell lines. In this study, we investigated the internalization mechanism by R10B/pDNA complex (R10B-lipoplex) in human prostate tumor PC-3 and human nasopharyngeal tumor KB cells, and compared with that in HeLa cells. Although it was necessary for R10B-lipoplex to associate with heparan sulfate (HS) on the cell surface in all cell lines, the R10B-lipoplex was internalized primarily through clathrin-mediated endocytosis in PC-3 and KB cells, and macropinosytosis in HeLa cells. In HeLa cells, treatment with the R10B-lipoplex induced the formation of lamellipodia for macropinocytosis, but did not in KB and PC-3 cells. Furthermore, the highest transfection efficiency by R10B-lipoplex was observed in HeLa cells. These findings indicated that the R10B-lipoplex induced the formation of lamellipodia in HeLa cells after binding to HS on the cells and was then internalized by macropinocytosis, which could induce high gene expression because of escaping degradation in lysosomes. Cell physiology might be a critical factor in cellular internalization and efficient transfection by cell penetration peptide.
authors.affiliation: Institute of Medicinal Chemistry, Hoshi University, Shinagawa-ku, Tokyo 142-8501, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Izumisawa
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Hattori
authors.forename: Masataka
authors.initials: M
authors.lastname: Date
authors.forename: Kazunori
authors.initials: K
authors.lastname: Toma
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Maitani
mesh_terms: D000199:Actins / Q000378:metabolism; D001120:Arginine / Q000031:analogs & derivatives / Q000737:chemistry*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D017930:Genes, Reporter; D006367:HeLa Cells; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D010455:Peptides / Q000737:chemistry*; D010873:Pinocytosis; D011092:Polyethylene Glycols / Q000737:chemistry*; D011554:Pseudopodia / Q000378:metabolism; D014162:Transfection
chemical_list: D000199:Actins; D001120:Arginine; D002966:Clathrin; D004247:DNA; D006497:Heparan Sulfate; D008055:Lipids; D008156:Luciferases; D010455:Peptides; D011092:Polyethylene Glycols; C015462:polyarginine