title: Apoptin Gene Delivery by the Functionalized Polyamidoamine Dendrimer Derivatives Induces Cell Death of U87-MG Glioblastoma Cells.
abstract: Malignant glioma is the most common and aggressive form of primary brain tumor in adults. In this study, we describe the efficacy of nonviral gene delivery carriers, histidine- and arginine- or histidine- and lysine-grafted polyamidoamine (PAMAM) dendrimers (PAMAM-H-R and PAMAM-H-K), in delivering a therapeutic and a tumor-selective killer gene, apoptin, using human glioma cells (U87-MG) and newborn human dermal fibroblast cells. We analyzed transfection efficiency using luciferase and a plasmid DNA encoding for enhanced green fluorescent protein and assessed cell viability in both cells. The results show that transfection efficiency of PAMAM-H-R and PAMAM-H-K was greatly increased compared with that of native PAMAM. Moreover, among PAMAM derivatives, cytotoxicity of PAMAM-H-K was very low. We treated both cells with complexes of PAMAM-H-R or PAMAM-H-K and apoptin and analyzed their cellular uptake by flow cytometry and localization by confocal microscopy. Furthermore, cell cycle distribution, caspase 3 activity assay, and JC-1 analysis showed cell death induced by apoptin in U87-MG cells. The present study demonstrates that a PAMAM-H-R/apoptin complex is an effective gene carrier system in glioma cell culture.
authors.affiliation: Department of Physiology, College of Medicine, Cardiovascular and Metabolic Disease Center, Inje University, Busan, Korea.
authors.forename: Yoonhee
authors.initials: Y
authors.lastname: Bae
authors.affiliation: Department of Biomedical Sciences, College of Medicine, The Catholic University of Korea, Seoul, Korea; Department of Medical Life Sciences, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Hyang-Shuk
authors.initials: HS
authors.lastname: Rhim
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon, Korea.
authors.forename: Seulgi
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Internal Medicine, Sanggye Paik Hospital, Cardiovascular and Metabolic Disease Center, Inje University, Seoul, Korea.
authors.forename: Kyung Soo
authors.initials: KS
authors.lastname: Ko
authors.affiliation: Department of Physiology, College of Medicine, Cardiovascular and Metabolic Disease Center, Inje University, Busan, Korea. Electronic address: phyhanj@inje.ac.kr.
authors.forename: Jin
authors.initials: J
authors.lastname: Han
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon, Korea. Electronic address: joonsig@cnu.ac.kr.
authors.forename: Joon Sig
authors.initials: JS
authors.lastname: Choi
mesh_terms: D017209:Apoptosis; D001120:Arginine / Q000031:analogs & derivatives; D001932:Brain Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D036022:Capsid Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D050091:Dendrimers / Q000737:chemistry*; D015316:Genetic Therapy / Q000379:methods; D005909:Glioblastoma / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D006639:Histidine / Q000031:analogs & derivatives; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives; D014162:Transfection / Q000379:methods*
chemical_list: D036022:Capsid Proteins; D050091:Dendrimers; C104700:PAMAM Starburst; C091717:VP3 protein, Chicken anemia virus; D006639:Histidine; D004247:DNA; D001120:Arginine; D008239:Lysine
keywords: U87-MG glioblastoma cells; apoptin; apoptosis; nonviral gene delivery