title: Dipeptide-functionalized polyamidoamine dendrimer-mediated apoptin gene delivery facilitates apoptosis of human primary glioma cells.
abstract: Glioblastoma multiform (GBM) is the most frequent and aggressive form of brain tumors in adults. However, the development of more efficient and safe nonviral vector gene therapy represents a promising therapeutic approach, using a tumor-specific killer gene, named apoptin. In this study, we describe the efficacy of non-viral gene delivery vectors, the amino acid-conjugated PAMAM derivatives (PAMAM-H-R and PAMAM-H-K) in delivering a therapeutic gene, displaying affinity toward human primary glioma cells (GBL-14 cells) and dermal fibroblasts. We analyzed transfection efficiency, using luciferase (Luci) and a pDNA encoding for enhanced fluorescent protein (EGFP), and cytotoxicity in both cells. The results show that transfection efficiency of PAMAM-H-R improved compared to native PAMAM dendrimer, but cytotoxicity of PAMAM-H-R and PAMAM-H-K were very low. We treated both cells with a polyplex formation of PAMAM-H-R or PAMAM-H-K/apoptin, and analyzed their cellular uptake and localization by flow cytometry and confocal microscopy. Furthermore, we analyzed the endosomal escape effect using TEM images, and found that PAMAM-H-R showed very fast escape from endosome to the cytosol. Caspase 3 activity assay, cell cycle distribution, and JC-1 analysis showed apoptosis induced by apoptin in GBL-14 cells. This indicates that PAMAM-H-R can be a potential nonviral vector gene delivery carrier for brain tumor therapy. The present study demonstrates that PAMAM-H-R/apoptin gene polyplex can be used as an effective therapeutic candidate for GBM due to its selective induction of apoptosis in primary glioma cells as a potential nonviral gene delivery carrier for brain tumor therapy.
authors.affiliation: Department of Physiology, College of Medicine, Cardiovascular and Metabolic Disease Center, Inje University, Busan 614-735, Republic of Korea.
authors.forename: Yoonhee
authors.initials: Y
authors.lastname: Bae
authors.affiliation: Salt Lake Community College, Salt Lake City, UT, USA.
authors.forename: Eric S
authors.initials: ES
authors.lastname: Green
authors.affiliation: Section on Cellular Differentiation, Program on Developmental Endocrinology and Genetics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Goo-Young
authors.initials: GY
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon 305-764, Republic of Korea.
authors.forename: Su Jeong
authors.initials: SJ
authors.lastname: Song
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Science, Eulji University, Republic of Korea.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Mun
authors.affiliation: Cell Engineering for Origin Research Center 46-21, Susong-dog, Jongno-gu, Seoul 110-140, Republic of Korea.
authors.forename: Sunray
authors.initials: S
authors.lastname: Lee
authors.affiliation: Chungnam National University School of Medicine, Daejeon 301-747, Republic of Korea.
authors.forename: Jong-Il
authors.initials: JI
authors.lastname: Park
authors.affiliation: School of Chemistry, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Jong-Sang
authors.initials: JS
authors.lastname: Park
authors.affiliation: Department of Internal Medicine, Sanggye Paik Hospital, Cardiovascular and Metabolic Disease Center, Inje University, Seoul 139-707, Republic of 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 614-735, Republic of 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 305-764, Republic of Korea. Electronic address: joonsig@cnu.ac.kr.
authors.forename: Joon Sig
authors.initials: JS
authors.lastname: Choi
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D050091:Dendrimers / Q000008:administration & dosage*; D004151:Dipeptides / Q000008:administration & dosage*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage; D005910:Glioma / Q000188:drug therapy*; D006801:Humans; D008156:Luciferases / Q000008:administration & dosage; D011073:Polyamines / Q000008:administration & dosage*; D014162:Transfection / Q000379:methods
chemical_list: D050091:Dendrimers; D004151:Dipeptides; C104700:PAMAM Starburst; C531249:Poly(amidoamine); D011073:Polyamines; D008156:Luciferases
keywords: Apoptin; Apoptosis; Gene therapy; Human primary glioma cells; PAMAM dendrimer