Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Jan 1;97(1):187–195. doi: 10.1172/JCI118388

Heparin-binding secretory transforming gene (hst) facilitates rat lactotrope cell tumorigenesis and induces prolactin gene transcription.

I Shimon 1, A Hüttner 1, J Said 1, O M Spirina 1, S Melmed 1
PMCID: PMC507078  PMID: 8550832

Abstract

We have shown previously that human prolactinomas express transforming sequences of the heparin-binding secretory transforming gene (hst) which encodes fibroblast growth factor-4 (FGF-4). To elucidate the role of hst in pituitary tumorigenesis we treated primary rat pituitary and pituitary tumor cell cultures with recombinant FGF-4 and also stably transfected pituitary cell lines with full-length human hst cDNA. Transfectants were screened for hst mRNA expression and FGF-4 production. FGF-4 (0.1-50 ng/ml) caused a dose-dependent 2.5-fold increase of prolactin (PRL) secretion (P < 0.001) in GH4 cells and up to 60% (P < 0.05) in primary cultures, while decreasing growth hormone release (P < 0.001). GH4 hst transfectants displayed markedly enhanced basal PRL secretion (threefold, P < 0.001) and also proliferated faster (P < 0.001). FGF-4 treatment of wild-type GH4 cells, transiently transfected with an expression construct (rPRL.luc) containing a luciferase reporter driven by the rPRL promoter, resulted in a dose-dependent increase of up to 3.3-fold in PRL transcriptional activity. Tumors derived from in vivo subcutaneous injection of GH4 hst-transfected cells strongly expressing FGF-4 grew more aggressively as assessed by histologic invasiveness and proliferating cell nuclear antigen staining (P < 0.01). The results indicate that hst overexpression mediates lactotrope tumor growth and potently stimulates PRL synthesis. Thus, hst may directly facilitate prolactinoma development via paracrine or autocrine action of its secreted protein, FGF-4.

Full Text

The Full Text of this article is available as a PDF (508.3 KB).

Selected References

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

  1. Adelaide J., Mattei M. G., Marics I., Raybaud F., Planche J., De Lapeyriere O., Birnbaum D. Chromosomal localization of the hst oncogene and its co-amplification with the int.2 oncogene in a human melanoma. Oncogene. 1988 Apr;2(4):413–416. [PubMed] [Google Scholar]
  2. Akita S., Webster J., Ren S. G., Takino H., Said J., Zand O., Melmed S. Human and murine pituitary expression of leukemia inhibitory factor. Novel intrapituitary regulation of adrenocorticotropin hormone synthesis and secretion. J Clin Invest. 1995 Mar;95(3):1288–1298. doi: 10.1172/JCI117779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alexander J. M., Biller B. M., Bikkal H., Zervas N. T., Arnold A., Klibanski A. Clinically nonfunctioning pituitary tumors are monoclonal in origin. J Clin Invest. 1990 Jul;86(1):336–340. doi: 10.1172/JCI114705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Atkin S. L., Landolt A. M., Jeffreys R. V., Diver M., Radcliffe J., White M. C. Basic fibroblastic growth factor stimulates prolactin secretion from human anterior pituitary adenomas without affecting adenoma cell proliferation. J Clin Endocrinol Metab. 1993 Sep;77(3):831–837. doi: 10.1210/jcem.77.3.8370706. [DOI] [PubMed] [Google Scholar]
  5. Bacus S., Flowers J. L., Press M. F., Bacus J. W., McCarty K. S., Jr The evaluation of estrogen receptor in primary breast carcinoma by computer-assisted image analysis. Am J Clin Pathol. 1988 Sep;90(3):233–239. doi: 10.1093/ajcp/90.3.233. [DOI] [PubMed] [Google Scholar]
  6. Baird A., Mormède P., Ying S. Y., Wehrenberg W. B., Ueno N., Ling N., Guillemin R. A nonmitogenic pituitary function of fibroblast growth factor: regulation of thyrotropin and prolactin secretion. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5545–5549. doi: 10.1073/pnas.82.16.5545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Basilico C., Newman K. M., Curatola A. M., Talarico D., Mansukhani A., Velcich A., Delli-Bovi P. Expression and activation of the K-fgf oncogene. Ann N Y Acad Sci. 1989;567:95–103. doi: 10.1111/j.1749-6632.1989.tb16462.x. [DOI] [PubMed] [Google Scholar]
  8. Benharroch D., Birnbaum D. Biology of the fibroblast growth factor gene family. Isr J Med Sci. 1990 Apr;26(4):212–219. [PubMed] [Google Scholar]
  9. Berwaer M., Monget P., Peers B., Mathy-Hartert M., Bellefroid E., Davis J. R., Belayew A., Martial J. A. Multihormonal regulation of the human prolactin gene expression from 5000 bp of its upstream sequence. Mol Cell Endocrinol. 1991 Sep;80(1-3):53–64. doi: 10.1016/0303-7207(91)90142-f. [DOI] [PubMed] [Google Scholar]
  10. Boggild M. D., Jenkinson S., Pistorello M., Boscaro M., Scanarini M., McTernan P., Perrett C. W., Thakker R. V., Clayton R. N. Molecular genetic studies of sporadic pituitary tumors. J Clin Endocrinol Metab. 1994 Feb;78(2):387–392. doi: 10.1210/jcem.78.2.8106627. [DOI] [PubMed] [Google Scholar]
  11. Brookes S., Smith R., Thurlow J., Dickson C., Peters G. The mouse homologue of hst/k-FGF: sequence, genome organization and location relative to int-2. Nucleic Acids Res. 1989 Jun 12;17(11):4037–4045. doi: 10.1093/nar/17.11.4037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Byström C., Larsson C., Blomberg C., Sandelin K., Falkmer U., Skogseid B., Oberg K., Werner S., Nordenskjöld M. Localization of the MEN1 gene to a small region within chromosome 11q13 by deletion mapping in tumors. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1968–1972. doi: 10.1073/pnas.87.5.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Böhlen P., Baird A., Esch F., Ling N., Gospodarowicz D. Isolation and partial molecular characterization of pituitary fibroblast growth factor. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5364–5368. doi: 10.1073/pnas.81.17.5364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  15. Delli Bovi P., Basilico C. Isolation of a rearranged human transforming gene following transfection of Kaposi sarcoma DNA. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5660–5664. doi: 10.1073/pnas.84.16.5660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Delli Bovi P., Curatola A. M., Kern F. G., Greco A., Ittmann M., Basilico C. An oncogene isolated by transfection of Kaposi's sarcoma DNA encodes a growth factor that is a member of the FGF family. Cell. 1987 Aug 28;50(5):729–737. doi: 10.1016/0092-8674(87)90331-x. [DOI] [PubMed] [Google Scholar]
  17. Delli-Bovi P., Curatola A. M., Newman K. M., Sato Y., Moscatelli D., Hewick R. M., Rifkin D. B., Basilico C. Processing, secretion, and biological properties of a novel growth factor of the fibroblast growth factor family with oncogenic potential. Mol Cell Biol. 1988 Jul;8(7):2933–2941. doi: 10.1128/mcb.8.7.2933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Feldman B., Poueymirou W., Papaioannou V. E., DeChiara T. M., Goldfarb M. Requirement of FGF-4 for postimplantation mouse development. Science. 1995 Jan 13;267(5195):246–249. doi: 10.1126/science.7809630. [DOI] [PubMed] [Google Scholar]
  19. Felgner P. L., Gadek T. R., Holm M., Roman R., Chan H. W., Wenz M., Northrop J. P., Ringold G. M., Danielsen M. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7413–7417. doi: 10.1073/pnas.84.21.7413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gonsky R., Herman V., Melmed S., Fagin J. Transforming DNA sequences present in human prolactin-secreting pituitary tumors. Mol Endocrinol. 1991 Nov;5(11):1687–1695. doi: 10.1210/mend-5-11-1687. [DOI] [PubMed] [Google Scholar]
  21. Gospodarowicz D. Purification of a fibroblast growth factor from bovine pituitary. J Biol Chem. 1975 Apr 10;250(7):2515–2520. [PubMed] [Google Scholar]
  22. Hall P. A., Levison D. A., Woods A. L., Yu C. C., Kellock D. B., Watkins J. A., Barnes D. M., Gillett C. E., Camplejohn R., Dover R. Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol. 1990 Dec;162(4):285–294. doi: 10.1002/path.1711620403. [DOI] [PubMed] [Google Scholar]
  23. Herman V., Drazin N. Z., Gonsky R., Melmed S. Molecular screening of pituitary adenomas for gene mutations and rearrangements. J Clin Endocrinol Metab. 1993 Jul;77(1):50–55. doi: 10.1210/jcem.77.1.8100831. [DOI] [PubMed] [Google Scholar]
  24. Herman V., Fagin J., Gonsky R., Kovacs K., Melmed S. Clonal origin of pituitary adenomas. J Clin Endocrinol Metab. 1990 Dec;71(6):1427–1433. doi: 10.1210/jcem-71-6-1427. [DOI] [PubMed] [Google Scholar]
  25. Kan M., Wang F., Xu J., Crabb J. W., Hou J., McKeehan W. L. An essential heparin-binding domain in the fibroblast growth factor receptor kinase. Science. 1993 Mar 26;259(5103):1918–1921. doi: 10.1126/science.8456318. [DOI] [PubMed] [Google Scholar]
  26. Li Y., Koga M., Kasayama S., Matsumoto K., Arita N., Hayakawa T., Sato B. Identification and characterization of high molecular weight forms of basic fibroblast growth factor in human pituitary adenomas. J Clin Endocrinol Metab. 1992 Dec;75(6):1436–1441. doi: 10.1210/jcem.75.6.1464644. [DOI] [PubMed] [Google Scholar]
  27. Mason I. J. The ins and outs of fibroblast growth factors. Cell. 1994 Aug 26;78(4):547–552. doi: 10.1016/0092-8674(94)90520-7. [DOI] [PubMed] [Google Scholar]
  28. Miyagawa K., Sakamoto H., Yoshida T., Yamashita Y., Mitsui Y., Furusawa M., Maeda S., Takaku F., Sugimura T., Terada M. hst-1 transforming protein: expression in silkworm cells and characterization as a novel heparin-binding growth factor. Oncogene. 1988 Oct;3(4):383–389. [PubMed] [Google Scholar]
  29. Moscatelli D. High and low affinity binding sites for basic fibroblast growth factor on cultured cells: absence of a role for low affinity binding in the stimulation of plasminogen activator production by bovine capillary endothelial cells. J Cell Physiol. 1987 Apr;131(1):123–130. doi: 10.1002/jcp.1041310118. [DOI] [PubMed] [Google Scholar]
  30. Nakagama H., Ohnishi S., Imawari M., Hirai H., Takaku F., Sakamoto H., Terada M., Nagao M., Sugimura T. Identification of transforming genes as hst in DNA samples from two human hepatocellular carcinomas. Jpn J Cancer Res. 1987 Jul;78(7):651–654. [PubMed] [Google Scholar]
  31. Neufeld G., Gospodarowicz D. The identification and partial characterization of the fibroblast growth factor receptor of baby hamster kidney cells. J Biol Chem. 1985 Nov 5;260(25):13860–13868. [PubMed] [Google Scholar]
  32. Niswander L., Jeffrey S., Martin G. R., Tickle C. A positive feedback loop coordinates growth and patterning in the vertebrate limb. Nature. 1994 Oct 13;371(6498):609–612. doi: 10.1038/371609a0. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Rifkin D. B., Moscatelli D. Recent developments in the cell biology of basic fibroblast growth factor. J Cell Biol. 1989 Jul;109(1):1–6. doi: 10.1083/jcb.109.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sakamoto H., Mori M., Taira M., Yoshida T., Matsukawa S., Shimizu K., Sekiguchi M., Terada M., Sugimura T. Transforming gene from human stomach cancers and a noncancerous portion of stomach mucosa. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3997–4001. doi: 10.1073/pnas.83.11.3997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schechter J., Goldsmith P., Wilson C., Weiner R. Morphological evidence for the presence of arteries in human prolactinomas. J Clin Endocrinol Metab. 1988 Oct;67(4):713–719. doi: 10.1210/jcem-67-4-713. [DOI] [PubMed] [Google Scholar]
  38. Schonbrunn A., Krasnoff M., Westendorf J. M., Tashjian A. H., Jr Epidermal growth factor and thyrotropin-releasing hormone act similarly on a clonal pituitary cell strain. Modulation of hormone production and inhbition of cell proliferation. J Cell Biol. 1980 Jun;85(3):786–797. doi: 10.1083/jcb.85.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sebo T. J., Roche P. C., Witzig T. E., Kurtin P. J. Proliferative activity in non-Hodgkin's lymphomas. A comparison of the bromodeoxyuridine labeling index with PCNA immunostaining and quantitative image analysis. Am J Clin Pathol. 1993 Jun;99(6):668–672. doi: 10.1093/ajcp/99.6.668. [DOI] [PubMed] [Google Scholar]
  40. Strohmeyer T., Peter S., Hartmann M., Munemitsu S., Ackermann R., Ullrich A., Slamon D. J. Expression of the hst-1 and c-kit protooncogenes in human testicular germ cell tumors. Cancer Res. 1991 Apr 1;51(7):1811–1816. [PubMed] [Google Scholar]
  41. Supowit S. C., Potter E., Evans R. M., Rosenfeld M. G. Polypeptide hormone regulation of gene transcription: specific 5' genomic sequences are required for epidermal growth factor and phorbol ester regulation of prolactin gene expression. Proc Natl Acad Sci U S A. 1984 May;81(10):2975–2979. doi: 10.1073/pnas.81.10.2975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Taira M., Yoshida T., Miyagawa K., Sakamoto H., Terada M., Sugimura T. cDNA sequence of human transforming gene hst and identification of the coding sequence required for transforming activity. Proc Natl Acad Sci U S A. 1987 May;84(9):2980–2984. doi: 10.1073/pnas.84.9.2980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Talarico D., Basilico C. The K-fgf/hst oncogene induces transformation through an autocrine mechanism that requires extracellular stimulation of the mitogenic pathway. Mol Cell Biol. 1991 Feb;11(2):1138–1145. doi: 10.1128/mcb.11.2.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tsuda T., Nakatani H., Matsumura T., Yoshida K., Tahara E., Nishihira T., Sakamoto H., Yoshida T., Terada M., Sugimura T. Amplification of the hst-1 gene in human esophageal carcinomas. Jpn J Cancer Res. 1988 May;79(5):584–588. doi: 10.1111/j.1349-7006.1988.tb00026.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Vallar L., Spada A., Giannattasio G. Altered Gs and adenylate cyclase activity in human GH-secreting pituitary adenomas. Nature. 1987 Dec 10;330(6148):566–568. doi: 10.1038/330566a0. [DOI] [PubMed] [Google Scholar]
  46. Velcich A., Delli-Bovi P., Mansukhani A., Ziff E. B., Basilico C. Expression of the K-fgf protooncogene is repressed during differentiation of F9 cells. Oncogene Res. 1989;5(1):31–37. [PubMed] [Google Scholar]
  47. Yoshida M. C., Wada M., Satoh H., Yoshida T., Sakamoto H., Miyagawa K., Yokota J., Koda T., Kakinuma M., Sugimura T. Human HST1 (HSTF1) gene maps to chromosome band 11q13 and coamplifies with the INT2 gene in human cancer. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4861–4864. doi: 10.1073/pnas.85.13.4861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Yoshida T., Ishimaru K., Sakamoto H., Yokota J., Hirohashi S., Igarashi K., Sudo K., Terada M. Angiogenic activity of the recombinant hst-1 protein. Cancer Lett. 1994 Aug 15;83(1-2):261–268. doi: 10.1016/0304-3835(94)90328-x. [DOI] [PubMed] [Google Scholar]
  49. Yoshida T., Miyagawa K., Odagiri H., Sakamoto H., Little P. F., Terada M., Sugimura T. Genomic sequence of hst, a transforming gene encoding a protein homologous to fibroblast growth factors and the int-2-encoded protein. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7305–7309. doi: 10.1073/pnas.84.20.7305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Zimering M. B., Brandi M. L., deGrange D. A., Marx S. J., Streeten E., Katsumata N., Murphy P. R., Sato Y., Friesen H. G., Aurbach G. D. Circulating fibroblast growth factor-like substance in familial multiple endocrine neoplasia type 1. J Clin Endocrinol Metab. 1990 Jan;70(1):149–154. doi: 10.1210/jcem-70-1-149. [DOI] [PubMed] [Google Scholar]
  51. Zimering M. B., Katsumata N., Sato Y., Brandi M. L., Aurbach G. D., Marx S. J., Friesen H. G. Increased basic fibroblast growth factor in plasma from multiple endocrine neoplasia type 1: relation to pituitary tumor. J Clin Endocrinol Metab. 1993 May;76(5):1182–1187. doi: 10.1210/jcem.76.5.8098714. [DOI] [PubMed] [Google Scholar]
  52. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES