Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells

Katsunori Yanagihara, Helen Cheng, Pi-Wan Cheng

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

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 (pCMVIacZ) using Lipofectin plus TF, insulin, or epidermal growth factor as the vector. The transfected cells were assessed for transfection efficiency by β-galactosidase activity (light units/μg protein) and the percentage of blue cells following 5-bromo- 4-chloro-3-indolyl β-D-galactopyranoside staining. Lipofectin supplemented with epidermal growth factor yielded the largest enhancement of lipofection efficiency (≤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.

Original languageEnglish (US)
Pages (from-to)59-65
Number of pages7
JournalCancer Gene Therapy
Volume7
Issue number1
DOIs
StatePublished - Jan 1 2000

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Transferrin
Epidermal Growth Factor
Insulin
Carcinoma
Transfection
Lung
Galactosidases
Ligands
Gene Targeting
DNA
Galactose
Reporter Genes
HeLa Cells
Plasmids
Staining and Labeling
Light
Cell Line
1,2-dielaidoylphosphatidylethanolamine
Genes
Proteins

Keywords

  • Gene therapy
  • Gene transfer
  • Ligand
  • Lipofection
  • Liposome
  • Nonviral vector

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Cancer Research

Cite this

Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells. / Yanagihara, Katsunori; Cheng, Helen; Cheng, Pi-Wan.

In: Cancer Gene Therapy, Vol. 7, No. 1, 01.01.2000, p. 59-65.

Research output: Contribution to journalArticle

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