Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Mar 15;91(6):2181–2185. doi: 10.1073/pnas.91.6.2181

Dominant negative inhibition of tumorigenesis in vivo by human insulin-like growth factor I receptor mutant.

D Prager 1, H L Li 1, S Asa 1, S Melmed 1
PMCID: PMC43334  PMID: 8134369

Abstract

Although insulin-like growth factor I (IGF-I) is a mitogenic growth factor, its role in tumorigenesis is unclear. We therefore transfected wild-type and truncated beta-subunit mutant (952STOP) human IGF-I receptor cDNAs into Rat-1 fibroblasts. Rat-1 transfectants expressed 2.5- to 7-fold increased IGF-I receptor mass, while the Kd for IGF-I binding was unchanged. The Rat-1 cells transfected with wild-type receptor cDNA responded to in vitro IGF-I treatment by increased proliferation and DNA synthesis. Cells overexpressing wild-type receptors were also transformed as evidenced by ligand-dependent colony proliferation in soft agar. After injection into athymic nude mice, all wild-type transfectants formed solid sarcomas within 3 weeks, and ex vivo tumor cell assays confirmed continued overexpression of human IGF-I receptors. In contrast, both DNA synthesis and proliferation of 952STOP-transfected cells were attenuated below that of untransfected cells. 952STOP cells were nonresponsive to IGF-I in vitro and were unable to sustain anchorage-independent growth. No tumors were induced for up to 8 weeks after injection of 952STOP transfectants into athymic mice, despite the presence of demonstrable endogenous IGF-I receptors on the 952STOP-transfected cells. Therefore, 952STOP behaves as a dominant negative inhibitor of endogenous IGF-I receptor function, probably by assembling nonfunctional hybrid rat/mutant human receptor tetramers.

Full text

PDF
2181

Images in this article

2182Image
on p.2182
2183Image
on p.2183

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Carpenter G. Receptor tyrosine kinase substrates: src homology domains and signal transduction. FASEB J. 1992 Nov;6(14):3283–3289. doi: 10.1096/fasebj.6.14.1385243. [DOI] [PubMed] [Google Scholar]
  2. Chatterjee V. K., Nagaya T., Madison L. D., Datta S., Rentoumis A., Jameson J. L. Thyroid hormone resistance syndrome. Inhibition of normal receptor function by mutant thyroid hormone receptors. J Clin Invest. 1991 Jun;87(6):1977–1984. doi: 10.1172/JCI115225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Daughaday W. H. The possible autocrine/paracrine and endocrine roles of insulin-like growth factors of human tumors. Endocrinology. 1990 Jul;127(1):1–4. doi: 10.1210/endo-127-1-1. [DOI] [PubMed] [Google Scholar]
  4. Downward J., Yarden Y., Mayes E., Scrace G., Totty N., Stockwell P., Ullrich A., Schlessinger J., Waterfield M. D. Close similarity of epidermal growth factor receptor and v-erb-B oncogene protein sequences. Nature. 1984 Feb 9;307(5951):521–527. doi: 10.1038/307521a0. [DOI] [PubMed] [Google Scholar]
  5. Drummond I. A., Madden S. L., Rohwer-Nutter P., Bell G. I., Sukhatme V. P., Rauscher F. J., 3rd Repression of the insulin-like growth factor II gene by the Wilms tumor suppressor WT1. Science. 1992 Jul 31;257(5070):674–678. doi: 10.1126/science.1323141. [DOI] [PubMed] [Google Scholar]
  6. Heldin C. H., Ernlund A., Rorsman C., Rönnstrand L. Dimerization of B-type platelet-derived growth factor receptors occurs after ligand binding and is closely associated with receptor kinase activation. J Biol Chem. 1989 May 25;264(15):8905–8912. [PubMed] [Google Scholar]
  7. Kaleko M., Rutter W. J., Miller A. D. Overexpression of the human insulinlike growth factor I receptor promotes ligand-dependent neoplastic transformation. Mol Cell Biol. 1990 Feb;10(2):464–473. doi: 10.1128/mcb.10.2.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kohn E. C., Francis E. A., Liotta L. A., Schiffmann E. Heterogeneity of the motility responses in malignant tumor cells: a biological basis for the diversity and homing of metastatic cells. Int J Cancer. 1990 Aug 15;46(2):287–292. doi: 10.1002/ijc.2910460225. [DOI] [PubMed] [Google Scholar]
  9. Macaulay V. M., Everard M. J., Teale J. D., Trott P. A., Van Wyk J. J., Smith I. E., Millar J. L. Autocrine function for insulin-like growth factor I in human small cell lung cancer cell lines and fresh tumor cells. Cancer Res. 1990 Apr 15;50(8):2511–2517. [PubMed] [Google Scholar]
  10. Marcelli M., Zoppi S., Grino P. B., Griffin J. E., Wilson J. D., McPhaul M. J. A mutation in the DNA-binding domain of the androgen receptor gene causes complete testicular feminization in a patient with receptor-positive androgen resistance. J Clin Invest. 1991 Mar;87(3):1123–1126. doi: 10.1172/JCI115076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Melmed S. Acromegaly. N Engl J Med. 1990 Apr 5;322(14):966–977. doi: 10.1056/NEJM199004053221405. [DOI] [PubMed] [Google Scholar]
  12. Okajima T., Nakamura K., Zhang H., Ling N., Tanabe T., Yasuda T., Rosenfeld R. G. Sensitive colorimetric bioassays for insulin-like growth factor (IGF) stimulation of cell proliferation and glucose consumption: use in studies of IGF analogs. Endocrinology. 1992 Apr;130(4):2201–2212. doi: 10.1210/endo.130.4.1372238. [DOI] [PubMed] [Google Scholar]
  13. Pendergast A. M., Muller A. J., Havlik M. H., Maru Y., Witte O. N. BCR sequences essential for transformation by the BCR-ABL oncogene bind to the ABL SH2 regulatory domain in a non-phosphotyrosine-dependent manner. Cell. 1991 Jul 12;66(1):161–171. doi: 10.1016/0092-8674(91)90148-r. [DOI] [PubMed] [Google Scholar]
  14. Pietrzkowski Z., Wernicke D., Porcu P., Jameson B. A., Baserga R. Inhibition of cellular proliferation by peptide analogues of insulin-like growth factor 1. Cancer Res. 1992 Dec 1;52(23):6447–6451. [PubMed] [Google Scholar]
  15. Poon B., Dixon D., Ellis L., Roth R. A., Rutter W. J., Wang L. H. Molecular basis of the activation of the tumorigenic potential of Gag-insulin receptor chimeras. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):877–881. doi: 10.1073/pnas.88.3.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Prager D., Yamasaki H., Weber M. M., Gebremedhin S., Melmed S. Human insulin-like growth factor I receptor function in pituitary cells is suppressed by a dominant negative mutant. J Clin Invest. 1992 Nov;90(5):2117–2122. doi: 10.1172/JCI116096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Redemann N., Holzmann B., von Rüden T., Wagner E. F., Schlessinger J., Ullrich A. Anti-oncogenic activity of signalling-defective epidermal growth factor receptor mutants. Mol Cell Biol. 1992 Feb;12(2):491–498. doi: 10.1128/mcb.12.2.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Roussel M. F., Dull T. J., Rettenmier C. W., Ralph P., Ullrich A., Sherr C. J. Transforming potential of the c-fms proto-oncogene (CSF-1 receptor). Nature. 1987 Feb 5;325(6104):549–552. doi: 10.1038/325549a0. [DOI] [PubMed] [Google Scholar]
  19. Schally A. V. Oncological applications of somatostatin analogues. Cancer Res. 1988 Dec 15;48(24 Pt 1):6977–6985. [PubMed] [Google Scholar]
  20. Sugarman B. J., Aggarwal B. B., Hass P. E., Figari I. S., Palladino M. A., Jr, Shepard H. M. Recombinant human tumor necrosis factor-alpha: effects on proliferation of normal and transformed cells in vitro. Science. 1985 Nov 22;230(4728):943–945. doi: 10.1126/science.3933111. [DOI] [PubMed] [Google Scholar]
  21. Taylor S. I., Cama A., Accili D., Barbetti F., Imano E., Kadowaki H., Kadowaki T. Genetic basis of endocrine disease. 1. Molecular genetics of insulin resistant diabetes mellitus. J Clin Endocrinol Metab. 1991 Dec;73(6):1158–1163. doi: 10.1210/jcem-73-6-1158. [DOI] [PubMed] [Google Scholar]
  22. Treadway J. L., Morrison B. D., Soos M. A., Siddle K., Olefsky J., Ullrich A., McClain D. A., Pessin J. E. Transdominant inhibition of tyrosine kinase activity in mutant insulin/insulin-like growth factor I hybrid receptors. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):214–218. doi: 10.1073/pnas.88.1.214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ueno H., Gunn M., Dell K., Tseng A., Jr, Williams L. A truncated form of fibroblast growth factor receptor 1 inhibits signal transduction by multiple types of fibroblast growth factor receptor. J Biol Chem. 1992 Jan 25;267(3):1470–1476. [PubMed] [Google Scholar]
  24. Velu T. J., Beguinot L., Vass W. C., Willingham M. C., Merlino G. T., Pastan I., Lowy D. R. Epidermal-growth-factor-dependent transformation by a human EGF receptor proto-oncogene. Science. 1987 Dec 4;238(4832):1408–1410. doi: 10.1126/science.3500513. [DOI] [PubMed] [Google Scholar]
  25. Waterfield M. D., Scrace G. T., Whittle N., Stroobant P., Johnsson A., Wasteson A., Westermark B., Heldin C. H., Huang J. S., Deuel T. F. Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus. Nature. 1983 Jul 7;304(5921):35–39. doi: 10.1038/304035a0. [DOI] [PubMed] [Google Scholar]
  26. Wilden P. A., Siddle K., Haring E., Backer J. M., White M. F., Kahn C. R. The role of insulin receptor kinase domain autophosphorylation in receptor-mediated activities. Analysis with insulin and anti-receptor antibodies. J Biol Chem. 1992 Jul 5;267(19):13719–13727. [PubMed] [Google Scholar]
  27. Yagi H., Ozono K., Miyake H., Nagashima K., Kuroume T., Pike J. W. A new point mutation in the deoxyribonucleic acid-binding domain of the vitamin D receptor in a kindred with hereditary 1,25-dihydroxyvitamin D-resistant rickets. J Clin Endocrinol Metab. 1993 Feb;76(2):509–512. doi: 10.1210/jcem.76.2.8381803. [DOI] [PubMed] [Google Scholar]
  28. Yamasaki H., Prager D., Melmed S. Structure-function of the human insulin-like growth factor-I receptor: a discordance of somatotroph internalization and signaling. Mol Endocrinol. 1993 May;7(5):681–685. doi: 10.1210/mend.7.5.8316251. [DOI] [PubMed] [Google Scholar]
  29. van Rijswijk R. E., van Loenen A. C., Wagstaff J., Meijer E., Lopez R., van Groeningen C. J., Heimans J. J., Pinedo H. M. Suramin: rapid loading and weekly maintenance regimens for cancer patients. J Clin Oncol. 1992 Nov;10(11):1788–1794. doi: 10.1200/JCO.1992.10.11.1788. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES