title: Transfection of lung cells in vitro and in vivo: effect of antioxidants and intraliposomal bFGF.
abstract: We hypothesized that constitutive formation of reactive oxygen species by respiratory cells is a barrier to gene transfer when liposome-DNA complexes are used, by contributing to rapid degradation of plasmid DNA. When plasmid DNA is complexed to liposomes it is protected against H(2)O(2)-mediated degradation but not that mediated by the hydroxyl radical. Treatment of distal rat fetal lung epithelial cells (RFL(19)Ep) with the vitamin E analog Trolox (50 microM) reduced intracellular plasmid degradation. Both Trolox (50 microM) and an iron chelator, phenanthroline (0.1 microM), significantly increased transgene expression in RFL(19)Ep approximately twofold, consistent with a hydroxyl radical-mediated inhibition of transgene expression. When basic fibroblast growth factor (bFGF; 20 ng/ml), a growth factor with antioxidant properties, was included within liposomes, we observed a significantly greater enhancement of RFL(19)Ep transgene expression (approximately 2-fold) over that seen with Trolox or phenanthroline. Inclusion of bFGF within liposomes also significantly enhanced (approximately 4-fold) transgene expression in mice following intratracheal instillation.
authors.affiliation: Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430074, People's Republic of China.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Luo
authors.forename: Rosetta
authors.initials: R
authors.lastname: Belcastro
authors.forename: Judy
authors.initials: J
authors.lastname: Cabacungan
authors.forename: Vicky
authors.initials: V
authors.lastname: Hannam
authors.forename: Anna
authors.initials: A
authors.lastname: Negus
authors.forename: Yanxia
authors.initials: Y
authors.lastname: Wen
authors.forename: Jonathan
authors.initials: J
authors.lastname: Plumb
authors.forename: Jim
authors.initials: J
authors.lastname: Hu
authors.forename: Brent
authors.initials: B
authors.lastname: Steer
authors.forename: David R
authors.initials: DR
authors.lastname: Koehler
authors.forename: Gregory P
authors.initials: GP
authors.lastname: Downey
authors.forename: A Keith
authors.initials: AK
authors.lastname: Tanswell
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D008081:Liposomes / Q000494:pharmacology; D008168:Lung / Q000166:cytology*; D016877:Oxidants / Q000494:pharmacology; D010957:Plasmids / Q000187:drug effects / Q000493:pharmacokinetics; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics
chemical_list: D000975:Antioxidants; D002839:Chromans; D008081:Liposomes; D016877:Oxidants; D017382:Reactive Oxygen Species; D016222:Fibroblast Growth Factor 2; D006861:Hydrogen Peroxide; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid