European Archives of Oto-Rhino-Laryngology

, Volume 254, Supplement 1, pp S130–S132 | Cite as

In vitro studies of liposome-mediated gene transfer into head and neck cancer cell lines

  • B. Wollenberg
  • S. Lang
  • B. Schmitt
  • E. Kastenbauer
  • R. Zeidler


The 5-year survival rate of patients with squamous cell carcinoma of the head and neck (HNSCC) has remained poor despite innovative surgery and new radiation and chemotherapeutic strategies. In such patients, gene therapy relying on the modification of tumor cells by gene transfer may have great potential as a new treatment modality in the therapy of HNSCC. In the present study we developed an in vitro model to show the efficacy and technical feasibility of cationic liposome-mediated gene transfer into HNSCC. Five adherent squamous cell carcinoma cell lines were transfected with SV40- or CMV promoter-driven CAT (chloramphenicol-acetyl-transferase)-expression plasmids using DOTAP as the liposome carrier. The level of CAT expression was shown to correlate directly with the amount of transfected DNA and could be measured by a CAT-enzyme-linked immunosorbent assay. The results of gene transfer by liposome-DNA complexes obtained for all cell lines showed a dose-dependent efficacy correlating to the amount of DOTAP employed. The data demonstrate the successful in vitro transfection of epithelial cell lines with DNA, suggesting its usefulness as a new tool for head and neck cancer therapy in vivo.

Key words

Gene therapy Head and neck squamous cell carcinoma Liposome carrier 


  1. 1.
    Bertling WM, Gareis M, Paspaleeva V, Zimmer A, Kreuter J, Numberg E, Harrer P (1991) Use of liposomes, viral capsids, and nanoparticles as DNA carriers. Biotechnol Appl Biochem 13: 390–405PubMedGoogle Scholar
  2. 2.
    Capaccioli S, Di Pasquale G, Mini E, Mazzei T, Quattrone A (1993) Cationic lipids improve antisense oligonucleotide uptake and prevent degradation in cultured cells and in human serum. Biochem Biophys Res Commun 197: 818–825PubMedCrossRefGoogle Scholar
  3. 3.
    Caplen NJ, Kinrade E, Sorgi F, Gao X, Gruenert D, Geddes D, Coutelle C, Huang L, Alton EWFW, Williamson R (1995) In vitro liposome-mediated DNA transfection of epithelial cell lines using the cationic liposome DC-Chol/DOPE. Gene Ther 2: 603–613PubMedGoogle Scholar
  4. 4.
    Cometta K, Morgan RA, Anderson WF (1991) Safety issues related to retroviral-mediated gene transfer in humans. Hum Gene Ther 2: 5–14CrossRefGoogle Scholar
  5. 5.
    Felgner PL, Ringold GM (1989) Cationic liposome-mediated transfection. Nature 337: 387–388PubMedCrossRefGoogle Scholar
  6. 6.
    Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA 84: 7413–7417PubMedCrossRefGoogle Scholar
  7. 7.
    Gao X, Huang L (1991) A novel cationic liposome reagent for efficient transfection of mammalian cells. Biochem Biophys Res Commun 179: 280–285PubMedCrossRefGoogle Scholar
  8. 8.
    Gorman CM, Moffat LF, Howard BH (1982) Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 2: 1044–1051PubMedGoogle Scholar
  9. 9.
    Kawai S, Nishizawa M (1984) New procedure for DNA transfection with polycation and dimethyl sulfoxide. Mol Cell Biol 4:1172–1174PubMedGoogle Scholar
  10. 10.
    Ledley FD (1994) Non-viral gene therapy. Curr Opin Biotechnol 5:626–636PubMedCrossRefGoogle Scholar
  11. 11.
    Legendre JY, Szoka FC (1992) Delivery of plasmid DNA into mammalian cell lines using pH-sensitive liposomes: comparison with cationic liposomes. Pharm Res 9: 1235–1242PubMedCrossRefGoogle Scholar
  12. 12.
    Loyter A, Scangos GA, Ruddle FH (1982) Mechanisms of DNA uptake by mammalian cells: fate of exogenously added DNA monitored by the use of fluorescent dyes. Proc Natl Acad Sci USA 79: 422–426PubMedCrossRefGoogle Scholar
  13. 13.
    Malone RW, Felgner PL, Verma IM (1989) Cationic liposomemediated RNA transfection. Proc Natl Acad Sci USA 86: 6077–6081PubMedCrossRefGoogle Scholar
  14. 14.
    Nabel GJ, Nabel EG, Yang ZY, Fox BA, Plautz GE, Gao X, Huang L, Shu S, Gordon D, Chang AE (1993) Direct gene transfer with DNA-liposome complexes in melanoma: expression, biologic activity, and lack of toxicity in humans. Proc Natl Acad Sci USA 90: 11307–11311PubMedCrossRefGoogle Scholar
  15. 15.
    Neumann E, Schaefer Ridder M, Wang Y, Hofschneider PH (1982) Gene transfer into mouse lymphoma cells by electroporation in high electric fields. EMBO J 1: 841–845PubMedGoogle Scholar
  16. 16.
    O'Malley BW Jr, Ledley FD (1993) Somatic gene therapy. Methods for the present and future. Arch Otolaryngol Head Neck Surg 119: 1100–1107PubMedGoogle Scholar
  17. 17.
    Rose JK, Buonocore L, Whitt MA (1991) A new cationic liposome reagent mediating nearly quantitative transfection of animal cells. Biotechniques 10: 520–525PubMedGoogle Scholar
  18. 18.
    Shillitoe EJ, Lapeyre JN, Adler Storthz K (1994) Gene therapy — its potential in the management of oral cancer. Eur J Cancer B Oral Oncol 30B: 143–154PubMedCrossRefGoogle Scholar
  19. 19.
    Vokes EE, Weichselbaum RR, Lippmann SM, Hong WK (1993) Head and neck cancer. N Engl J Med 328: 184–191PubMedCrossRefGoogle Scholar
  20. 20.
    Watanabe Y, Nomoto H, Takezawa R, Miyoshi N, Akaike T (1994) Highly efficient transfection into primary cultured mouse hepatocytes by use of cation-liposomes: an application for immunization. J Biochem (Tokyo) 116: 1220–1226Google Scholar
  21. 21.
    Wattiaux R, Jadot M, Dubois F, Misquith S, Wattiaux De Coninck S (1995) Uptake of exogenous DNA by rat liver: effect of cationic lipids. Biochem Biophys Res Commun 213: 81–87PubMedCrossRefGoogle Scholar
  22. 22.
    Wollenberg B, Naujoks K, Kastenbauer E (1996) Somatische Gentherapie von Plattenepithelkarzinomen des Kopf-Hals-Bereiches. Laryngorhinootologie 75: 356–362PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • B. Wollenberg
    • 1
  • S. Lang
    • 1
  • B. Schmitt
    • 1
  • E. Kastenbauer
    • 1
  • R. Zeidler
    • 1
  1. 1.Department of Oto-Rhino-Laryngology, University of MunichGroßhadern Medical CenterMunichGermany

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