Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells.
Poor transfection efficiency is the major drawback of lipofection. We showed previously that addition of transferrin (TF) to Lipofectin enhanced the expression of a reporter gene in HeLa cells by 120-fold and achieved close to 100% transfection efficiency. The purpose of this study was to determine whether TF and other ligands could improve the efficiency of lipofection in lung carcinoma cells. Confluent A549, Calu3, and H292 cells were transfected for 18 hours with a plasmid DNA (pCMVlacZ) using Lipofectin plus TF, insulin, or epidermal growth factor as the vector. The transfected cells were assessed for transfection efficiency by beta-galactosidase activity (light units/microg protein) and the percentage of blue cells following 5-bromo-4-chloro-3-indolyl beta-D-galactopyranoside staining. Lipofectin supplemented with epidermal growth factor yielded the largest enhancement of lipofection efficiency (< or =23-fold over that by Lipofectin alone) in all three cell lines. Insulin significantly enhanced the lipofection efficiency in A549 and Calu3 cells but not in H292 cells, whereas TF showed significant lipofection efficiency-enhancing effect in Calu3 and H292 cells but not in A549 cells. The transfection efficiency correlated well with the amounts of DNA delivered to the nucleus as well as the amounts of the receptor. These results indicate that the gene delivery strategy employing ligand-facilitated lipofection can achieve high transfection efficiency in human lung carcinoma cells. In addition, enhancement of the expression of the receptor may be a possible strategy for increasing the efficiency of gene targeting.
Yanagihara
K
K
Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha 68198-4525, USA.
Cheng
H
H
Cheng
P W
PW
D002467:Cell Nucleus / Q000235:genetics; D004247:DNA / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D008024:Ligands; D008081:Liposomes; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D014162:Transfection / Q000379:methods*; D014168:Transferrin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D007328:Insulin; D008024:Ligands; D008081:Liposomes; D014168:Transferrin; D004815:Epidermal Growth Factor; D004247:DNA