Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Acquired Diseases
  • Published:

TGF-α antisense gene therapy inhibits head and neck squamous cell carcinoma growth in vivo

Gene Therapy volume 7, pages 1906–1914 (2000)Cite this article

Abstract

Unlike normal mucosal squamous epithelial cells, head and neck squamous cell carcinomas (HNSCCs) overexpress TGF-α mRNA and protein which is required to sustain the proliferation of HNSCC cells in vitro. To determine whether TGF-α expression contributes to tumor growth in vivo, cationic liposome-mediated gene transfer was used to deliver an antisense expression construct targeting the human TGF-α gene into human head and neck tumor cells, grown as subcutaneous xenografts in nude mice. The TGF-α antisense gene was immediately detected in the cytoplasm of the tumor cells, translocated to the nucleus by 12 h and remained localized to the nucleus for up to 3 days. Direct inoculation of the TGF-α antisense (but not the corresponding sense) construct into established HNSCC tumors resulted in inhibition of tumor growth. Sustained antitumor effects were observed for up to 1 year after the treatments were discontinued. Down-modulation of TGF-α was accompanied by increased apoptosis in vivo. These experiments indicate that interference with the TGF-α/EGFR autocrine signaling pathway may be an effective therapeutic strategy for cancers which overexpress this ligand/receptor pair.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Todaro GJ, De Larco JE . Transformation by murine and feline sarcoma viruses specifically blocks binding of epidermal growth factor to cells Nature 1976 264: 26–31

    Article  CAS  PubMed  Google Scholar 

  2. Todaro GJ, Fryling C, De Larco JE . Transforming growth factors produced by certain human tumor cells: polypeptides that interact with epidermal growth factor receptors Proc Natl Acad Sci USA 1980 77: 5258–5262

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Barrandon Y, Green H . Cell migration is essential for sustained growth of deratinocyte colonies: the roles of transforming growth factor-alpha and epidermal growth factor Cell 1987 50: 1131–1137

    Article  CAS  PubMed  Google Scholar 

  4. Coffey RJ Jr et al. Production and auto-induction of transforming growth factor-alpha in human keratinocytes Nature 1987 328: 817–820

    Article  PubMed  Google Scholar 

  5. Liscia DS et al. Transforming growth factor-alpha messenger RNA localization in the developing adult rat and human mammary gland by in situ hybridization Dev Biol 1990 140: 123–131

    Article  CAS  PubMed  Google Scholar 

  6. Thomas DM, Nasim MM, Gullick WJ, Alison MR . Immunoreactivity of transforming growth factor alpha in the normal adult gastrointestinal tract Gut 1992 33: 628–631

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Kudlow JE et al. Transforming growth factor-alpha in the mammalian brain. Immunohistochemical detection in neurons and characterization if its mRNA J Biol Chem 1989 264: 3880–3883

    CAS  PubMed  Google Scholar 

  8. Xynos FP et al. Expression of transforming growth factor alpha mRNA in benign and malignant tissues derived from gynecologic patients with various proliferative conditions Anticancer Res 1992 12: 1115–1120

    CAS  PubMed  Google Scholar 

  9. Gomella LG et al. Expression of transforming growth factor alpha in normal human adult kidney and enhanced expression of transforming growth factors alpha and beta 1 in renal cell carcinoma Cancer Res 1989 49: 6972–6975

    CAS  PubMed  Google Scholar 

  10. Rosenthal A et al. Expression in rat fibroblasts of a human transforming growth factor-alpha cDNA results in transformation Cell 1986 46: 301–309

    Article  CAS  PubMed  Google Scholar 

  11. Sandgren EP et al. Overexpression of TGF alpha in transgenic mice: induction of epithelial hyperplasia, pancreatic metaplasia, and carcinoma of the breast Cell 1990 61: 1121–1135

    Article  CAS  PubMed  Google Scholar 

  12. Gewirtz AM . Am Society of Clin Oncol-Educational Book Bostrom Corp: Chicago 1995 pp 192–196

    Google Scholar 

  13. Saxena A, Ali IU . Increased expression of genes from growth factor signaling pathways in glioblastoma cell lines Oncogene 1992 7: 243–247

    CAS  PubMed  Google Scholar 

  14. Bates SE, Davidson NE, Valverius EM . Expression of transforming growth factor alpha and its messenger ribonucleic acid in human breast cancer: its regulation by estrogen and its possible functional significance Mol Endocrinol 1988 2: 543–545

    Article  CAS  PubMed  Google Scholar 

  15. Dickson RB et al. Growth factors in breast cancer: mitogenesis to transformation J Steroid Biochem Mol Biol 1992 43: 69–78

    Article  CAS  PubMed  Google Scholar 

  16. Iihara K et al. Prognostic significance of transforming growth factor-alpha in human esophageal carcinoma. Implication for the autocrine proliferation Cancer 1993 71: 2902–2909

    Article  CAS  PubMed  Google Scholar 

  17. Smith JM, Derynck R, Korc M . Production of transforming growth factor alpha in human pancreatic cancer cells: evidence for a superagonist autocrine cycle Proc Natl Acad Sci USA 1987 84: 7567–7570

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Barton CM et al. Transforming growth factor alpha and epidermal growth factor in human pancreatic cancer J Pathol 1991 163: 111–116

    Article  CAS  PubMed  Google Scholar 

  19. Yeh YC et al. Elevation of transforming growth factor alpha and its relationship to the epidermal growth factor and alpha-fetoprotein levels in patients with hepatocellular carcinoma Cancer Res 1987 47: 896–901

    CAS  PubMed  Google Scholar 

  20. Liu C, Woo A, Tsao MS . Expression of transforming growth factor-alpha in primary human colon and lung carcinomas Br J Cancer 1990 62: 425–429

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Tanaka S et al. Immunoreactive transforming growth factor alpha is commonly present in colorectal neoplasia Am J Pathol 1991 139: 123–129

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Tateishi M et al. Immunohistochemical evidence of autocrine growth factors in adenocarcinoma of the human lung Cancer Res 1990 50: 7077–7080

    CAS  PubMed  Google Scholar 

  23. Mydlo JH et al. Expression of transforming growth factor alpha and epidermal growth factor receptor messenger RNA in neoplastic and nonneoplastic human kidney tissue Cancer Res 1989 49: 3407–3411

    CAS  PubMed  Google Scholar 

  24. Mellon JK, Cook S, Chambers P, Neal DE . Transforming growth factor alpha and epidermal growth factor levels in bladder cancer and their relationship to epidermal growth factor receptor Br J Cancer 1996 73: 654–658

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Reiss M, Stash EB, Vellucci VF, Zhou Z . Activation of the autocrine transforming growth factor alpha pathway in human squamous carcinoma cells Cancer Res 1991 51: 6254–6262

    CAS  PubMed  Google Scholar 

  26. Morishige K et al. Involvement of transforming growth factor alpha/epidermal growth factor receptor autocrine growth mechanism in an ovarian cancer cell line in vitro Cancer Res 1991 51: 5951–5955

    CAS  PubMed  Google Scholar 

  27. Todd R et al. TGF-alpha and EGF-receptor mRNAs in human oral cancers Carcinogenesis 1989 10: 1553–1556

    Article  CAS  PubMed  Google Scholar 

  28. Rubin Grandis J, Tweardy DJ . Elevated levels of transforming growth factor alpha and epidermal growth factor receptor messenger RNA are early markers of carcinogenesis in head and neck cancer Cancer Res 1993 53: 3579–3584

    Google Scholar 

  29. Rubin Grandis J, Melhem MF, Barnes EL, Tweardy DJ . Quantitative immunohistochemical analysis of transforming growth factor-alpha and epidermal growth factor receptor in patients with squamous cell carcinoma of the head and necck Cancer 1996 78: 1284–1292

    Article  CAS  PubMed  Google Scholar 

  30. Di Marco E et al. Autocrine interaction between TGF alpha and the EGF-receptor: quantitative requirements for induction of the malignant phenotype Oncogene 1989 4: 831–838

    CAS  PubMed  Google Scholar 

  31. Moroni MC, Willingham MC, Beguinot L . EGF-R antisense RNA blocks expression of the epidermal growth factor receptor and suppresses the transforming phenotype of a human carcinoma cell line J Biol Chem 1992 267: 2714–2722

    CAS  PubMed  Google Scholar 

  32. Atlas I et al. Growth regulation of human renal carcinoma cells: role of transforming growth factor alpha Cancer Res 1992 52: 335–339

    Google Scholar 

  33. Rubin Grandis J et al. Tumor levels of TGF-α and EGFR protein predict survival in patients with head and neck squamous cell carcimona J Natl Cancer Inst 1998 90: 824–832

    Article  CAS  PubMed  Google Scholar 

  34. Rubin Grandis J et al. Downmodulation of TGF-alpha protein expression with antisense oligonucleotides inhibits proliferation of head and neck squamous carcinoma but not normal mucosal epithelial cells J Cell Biochem 1998 69: 55–62

    Article  Google Scholar 

  35. He Y et al. Inhibition of human squamous cell carcinoma growth in vivo by epidermal growth factor receptor antisense RNA transcribed from the U6 promoter J Natl Cancer Inst 1998 90: 1080–1087

    Article  CAS  PubMed  Google Scholar 

  36. Rubin Grandis J, Zeng Q, Tweardy DJ . Retinoic acid normalizes the increased gene transcription rate of TGF-alpha and EGFR in head and neck cancer cell lines Nature Med 1996 2: 237–240

    Article  CAS  PubMed  Google Scholar 

  37. Rubin Grandis J, Tweardy DJ, Melhem MF . Asynchronous modulation of transforming growth factor-alpha and epidermal growth factor receptor protein expression in progression of premalignant lesions to head and neck squamous cell carcinoma Clin Cancer Res 1998 4: 13–20

    CAS  PubMed  Google Scholar 

  38. Imanishi KI et al. Inhibition of growth of human lung adenocarcinoma cell lines by anti-transforming growth factor-alpha monoclonal antibody J Natl Cancer Inst 1989 81: 220–223

    Article  CAS  PubMed  Google Scholar 

  39. Filmus J, Shi W, Spencer T . Role of transforming growth factor alpha (TGF-alpha) in the transformation of ras-transfected rat intestinal epithelial cells Oncogene 1993 8: 1017–1022

    CAS  PubMed  Google Scholar 

  40. Sizeland AW, Burgess AW . Antisense transforming growth factor alpha oligonucleotides inhibit autocrine stimulated proliferation of a colon carcinoma cell line Mol Biol Cell 1992 3: 1235–1243

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Solorzano CC et al. Inhibition of transforming growth factor alpha stimulation of human squamous cell carcinoma of the head and neck with anti-TGF-alpha antibodies and tyrphostin Ann Surg Oncol 1997 4: 670–678

    Article  CAS  PubMed  Google Scholar 

  42. Laird AD, Brown PI, Fausto N . Inhibition of tumor growth in liver epithelial cells transfected with a transforming growth factor alpha antisense gene Cancer Res 1994 54: 4224–4232

    CAS  PubMed  Google Scholar 

  43. Rubenstein M, Mirochnik Y, Chou P, Guinan P . Antisense oligonucleotide intralesional therapy for human PC-3 prostate tumors carried in athymic nude mice J Surg Oncol 1996 62: 194–200

    Article  CAS  PubMed  Google Scholar 

  44. Gewirtz AM et al. Bone marrow purging with oligodeoxynucleotides Blood 1996 88: 1517

    CAS  PubMed  Google Scholar 

  45. Webb A et al. Bcl-2 antisense therapy in patients with non-Hodgkin lymphoma Lancet 1997 349: 1137–1141

    Article  CAS  PubMed  Google Scholar 

  46. Cammilleri S et al. Biodistribution of iodine-125 tyramine transforming growth factor alpha antisense oligonucleotide in athymic mice with a human mammary tumor xenograft following intratumoral injection Eur J Nucl Med 1996 23: 448–452

    Article  CAS  PubMed  Google Scholar 

  47. He Y, Huang L . Growth inhibition of human papillomarvirus 16 DNA-positive mouse tumor by antisense RNA transcribed from U6 promoter Cancer Res 1997 57: 3993–3999

    CAS  PubMed  Google Scholar 

  48. Nabel GJ et al. Direct gene transfer with DNA–liposome complexes in melanoma: expression, biologic activity, and lack of toxicity in humans Proc Natl Acad Sci USA 1993 90: 11307–11311

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Caplen NJ et al. Liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis Nature Med 1995 1: 39–46

    Article  CAS  PubMed  Google Scholar 

  50. Noonberg SB et al. In vivo generation of highly abundant sequence-specific oligonucleotides for antisense and triplex gene regulation Nucleic Acids Res 1994 22: 2830–2836

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Mahr S et al. Growth factor effects on apoptosis of rat gastric enterochromaffin-like cells Endocrinology 1998 139: 4380–4390

    Article  CAS  PubMed  Google Scholar 

  52. Brison DR, Schultz RM . Increased incidence of apoptosis in transforming growth factor alpha-deficient mouse blastocytes Biol Reprod 1998 59: 136–144

    Article  CAS  PubMed  Google Scholar 

  53. Amundadottir LT et al. Cooperation of TGF alpha and c-myc in mouse mammary tumorigenesis: coordinated stimulation of growth and suppression of apoptosis Oncogene 1996 13: 757–765

    CAS  PubMed  Google Scholar 

  54. Seki S et al. Induction of apoptosis in a human hepatocellular carcinoma cell line by a neutralizing antibody to transforming growth factor-alpha Virchows Arch 1997 430: 29–35

    Article  CAS  PubMed  Google Scholar 

  55. Nakamura N, Shikoji Y, Moriwaki H, Muto Y . Apoptosis in human hempatoma cell line induced by 4,5-didehydro geranylgeranoic acid (acyclic acid) via down-regulation of transforming growth factor-alpha Biochem Biophys Res Commun 1996 219: 100–104

    Article  CAS  PubMed  Google Scholar 

  56. Rubin Grandis J, Chakraborty A, Melhem MF, Tweardy DJ . Inhibition of epidermal growth factor receptor gene expression and function decreases proliferation of head and neck squamous carcinoma but not normal mucosal epithelial cells Oncogene 1997 15: 409–416

    Article  CAS  PubMed  Google Scholar 

  57. Ciardiello F et al. Cooperative inhibition of renal cancer growth by anti-epidermal growth factor receptor antibody and protein kinase A antisense oligonucleotide J Natl Cancer Inst 1998 90: 1087–1094

    Article  CAS  PubMed  Google Scholar 

  58. De Luca A et al. Antisense oligonucleotides directed against EGF-related growth factors enhance anti-proliferative effect of conventional anti-tumor drugs in human colon-cancer cells Int J Cancer 1997 73: 277–282

    Article  CAS  PubMed  Google Scholar 

  59. Qian JF, Lazar-Wesley E, Breugnot C, May E . Human transforming growth factor alpha: sequence analysis of the 4.5-kb and 1.6-kb mRNA species Gene 1993 132: 291–296

    Article  CAS  PubMed  Google Scholar 

  60. Sacks PG et al. Establishment and characterization of two new squamous cell carcinoma cell lines derived from tumors of the head and neck Cancer Res 1988 48: 2858–2866

    CAS  PubMed  Google Scholar 

  61. Drenning SD et al. Bcl-2 but not Bax expression is associated with apoptosis in normal and transformed squamous epithelium Clin Cancer Res 1998 4: 2913–2921

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr Reuben Lotan for the gift of the cell line 1483. This work was supported in part by Public Health Service grants CA01760, CA72526, and CA77038 from The National Cancer Institute (to JRG), The Eye and Ear Foundation, The Pittsburgh Foundation, The Mary Hillman Jennings Foundation and the Veterans Research Foundation of Pittsburgh.

Author information

Authors and Affiliations

  1. Department of Otolaryngology, University of Pittsburgh School of Medicine, and University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA

    S Endo, Q Zeng, M N Lango, S D Drenning & J Rubin Grandis

  2. Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, and University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA

    N A Burke, S F Watkins & J Rubin Grandis

  3. Department of Pharmacology, University of Pittsburgh School of Medicine, and University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA

    Y He & L Huang

  4. Department of Pathology, University of Pittsburgh School of Medicine, and University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA

    M F Melhem

Authors
  1. S Endo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Q Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N A Burke
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Y He
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M F Melhem
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S F Watkins
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. M N Lango
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. S D Drenning
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. L Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. J Rubin Grandis
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Endo, S., Zeng, Q., Burke, N. et al. TGF-α antisense gene therapy inhibits head and neck squamous cell carcinoma growth in vivo. Gene Ther 7, 1906–1914 (2000). https://doi.org/10.1038/sj.gt.3301315

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301315

Keywords

This article is cited by

Search

Advanced search

Quick links