Transfection of lung cells in vitro and in vivo: effect of antioxidants and intraliposomal bFGF.
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.
Luo
Xiaoping
X
Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430074, People's Republic of China.
Belcastro
Rosetta
R
Cabacungan
Judy
J
Hannam
Vicky
V
Negus
Anna
A
Wen
Yanxia
Y
Plumb
Jonathan
J
Hu
Jim
J
Steer
Brent
B
Koehler
David R
DR
Downey
Gregory P
GP
Tanswell
A Keith
AK
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
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