Journal of Endocrinological Investigation

, Volume 18, Issue 2, pp 83–90 | Cite as

Interleukin-6 stimulates cell proliferation of rat pituitary clonal cell lines in vitro

  • T. Sawada
  • Koji Koike
  • Y. Kanda
  • H. Ikegami
  • H. Jikihara
  • T. Maeda
  • Y. Osako
  • K. Hirota
  • A. Miyake


We investigated the effect of recombinant human IL-6 (rhlL-6) on cell proliferation using the MtT/E rat pituitary tumor cell line, which was recently established by Inoue et al. This cell line expresses the homeodomain protein Pit-1/GHF 1 and does not produce any significant amount of pituitary hormones, but retains its tumorigenicity by back-transplantation into rats, resulting in production of prolactin. MtT/E cells were seeded into Falcon 24-well plates at a density of 2×104 cells/well in a cultured medium, containing 10% horse serum and 2.5% fetal bovine, with test drug. After four-days (12 days for the time-course study) incubations, the cells were counted using a hemocytometer. Incubation for 4 days with rhlL-6 caused concentration-dependent stimulation of MtT/E cell growth and [3H]-thymidine incorporation into MtT/E cells. Addition of 20 ng/ml rhlL-6 to the culture medium stimulated MtT/E cell growth in a time-dependent manner, withdrawal of rhlL-6 from the culture medium reduced MtT/E cell growth, and re-addition of rhlL-6 to the culture medium again stimulated MtT/E cell growth. Among the cytokines tested, granulocyte colony-stimulating factor (rh G-CSF) also showed a slight but significant mitogenic activity on the MtT/E cells. Analysis of 125|-rhlL-6 binding to the MtT/E cells indicated a dissociation constant of 0.953×10−9 mmol/l and the presence of 968 binding sites per cell. These results confirm the previous finding that IL-6 exerts mitogenic activity on pituitary tumor cells and also support the hypothsis that IL-6 may be involved in the control of cell growth and proliferation in the pituitary.


Interleukin-6 interleukin-1 pituitary tumor cell cell growth lnterleukin-6 binding site 


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  1. 1.
    Bazan J.F. A novel family of growth factor receptors: A common binding domain in the growth hormone, prolactin, erythropoietin and IL-6 receptors, and the p75 IL-2 receptor ß-chain. Biochem. Biophys. Res. Commun. 164: 788, 1989.PubMedCrossRefGoogle Scholar
  2. 2.
    Bernton E.W., Beach J.E., Holaday J.W., Smallridge R.C., Fein H.G. Release of multiple hormones by a direct action of interleukin-1 on pituitary cells. Science 238: 519, 1987.PubMedCrossRefGoogle Scholar
  3. 3.
    Spangelo B.L., Judd A.M., Isakson P.C., MacLeod R.M. lnterleukin-6 stimulates anterior pituitary hormone release in vitro. Endocrinology 125: 575, 1989.PubMedCrossRefGoogle Scholar
  4. 4.
    Naitoh Y., Fukuta J., Tominaga T., Nakai Y., Tamai S., Mori K., Imura H. lnterleukin-6 stimulates the secretion of adrenocorticotropic hormone in conscious, freely-moving rats. Biochem. Biophys. Res. Commun. 155: 1459, 1988.PubMedCrossRefGoogle Scholar
  5. 5.
    Beach J.E., Smallridge R.C., Kinzer C.A., Bernton E.W., Holaday J.W., Fein H.G. Rapid release of multiple hormones from rat pituitaries perifused with recombinant interleukin-1. Life Sci. 44: 1, 1989.PubMedCrossRefGoogle Scholar
  6. 6.
    Yamaguchi M., Matsuzaki N., Hirota K., Miyake A., Tanizawa O. lnterleukin-6 possibly induced by interleukin-1ß in the pituitary gland stimulates the releases of gonadotropins and prolactin. Acta Endocrinol. (Copenh.) 122: 201, 1990.Google Scholar
  7. 7.
    Jones T.H., Justice S., Price A., Chapman K. Lnterleukin-6 secreting human pituitary adenomas in vitro. J. Clin. Endocrinol. Metab. 73: 207, 1991.PubMedCrossRefGoogle Scholar
  8. 8.
    Jones T.H., Daniels M., James R.A., Justice S.K., McCorkle R., Price A., Kendall-Taylor P., Weetman A.P. Production of bioactive and immunoreactive inter-leukin-6 (IL-6) and expression of IL-6 messenger ribonucleic acid by human pituitary adenomas. J. Clin. Endocrinol. Metab. 78: 180, 1994.PubMedGoogle Scholar
  9. 9.
    Vankelecom H., Carmeliet P., Damme J.V., Billiau A., Denef C. Production of interleukin-6 by folliculo-stellate cells of the anterior pituitary gland in a histiotypic cell aggregate culture system. Neuroendocrinology 49: 102, 1989.PubMedCrossRefGoogle Scholar
  10. 10.
    Spangelo B.L., Judd A.M., Isakson P.C., MacLeod R.M. Interleukin-1 stimulates interleukin-6 release from rat anterior pituitary cells in vitro. Endocrinology 123: 2685, 1991.CrossRefGoogle Scholar
  11. 11.
    Tatsuno I., Somogyvari-vigh A., Mizuno K., Gottschall P.E., Hidaka H., Arimura A. Neuropeptide regulation of interleukin-6 production from the pituitary: Stimulation by pituitary adenylate cyclase activating polypeptide and calcitonin generelated peptide. Endocrinology 129: 1797, 1991.PubMedCrossRefGoogle Scholar
  12. 12.
    Ikebuchi K., Wong G.G., Clark S.C., Ihle J.N., Hirai Y., Ogawa M. Interleukin 6 enhancement of interleukin 3-dependent proliferation of multipotential hemopoietic progenitors. Proc. Natl. Acad. Sci. USA 84: 9035, 1987.PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Suematsu S., Matsuda T., Aozasa K., Akira S., Nakano N., Ohno S., Miyazaki J., Yamamura K., Hirano T., Kishimoto T. IgG 1 plasmacytosis in interleukin 6 transgenic mice. Proc. Natl. Acad. Sci. USA 86: 7547, 1989.PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Hirano T. Interleukin 6 (IL-6) and its receptor: Their role in plasma cell neoplasias. Int. J. Cell. Cloning 9: 166, 1991.PubMedCrossRefGoogle Scholar
  15. 15.
    Kawano M., Hirano T., Matsuda T., Taga T., Horii Y., Iwato K., Asaoku H., Tang B., Tanabe O., Tanaka H., Kuramoto A., Kishimoto T. Autocrine generation and requirement of BSF-2/IL-6 for human multiple myelomas. Nature (London) 332: 83, 1988.CrossRefGoogle Scholar
  16. 16.
    Koo A.S., Armstrong C., Bochner B., Shimabukuro T., Tso C., deKernion J.B., Belldegrun A. lnterleukin-6 and renal cell cancer: Production, regulation, and growth effects. Cancer Immunol. Immunother. 35: 97, 1992.PubMedCrossRefGoogle Scholar
  17. 17.
    May L.T., Torcia G., Cozzolino F., Ray A., Tatter S.B., Santhanam V., Sehgal P.B., Stern D. lnterleukin-6 gene expression in human endothelial cells: RNA start sites, multiple IL-6 proteins and inhibition of proliferation. Biochem. Biophys. Res. Commun. 759: 991, 1989.CrossRefGoogle Scholar
  18. 18.
    Chen L, Mory Y., Zilberstein A., Rovel M. Growth inhibition of human breast carcinoma and leukemia/lymphoma cell lines by recombinant interferon-ß2. Proc. Natl. Acad. Sci. USA 85: 8037, 1988.PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Tamm I., Cardinale I., Krueger J., Mupphy J.S., May L.T., Sehgal P.B. Interleukin 6 decreases cell-cell association and increases motility of ductal breast carcinoma cells. J. Exp. Med. 170: 1649, 1989.PubMedCrossRefGoogle Scholar
  20. 20.
    Arzt E., Buric R, Stelzer G., Stalla J., Sauer J., Renner U., Stalla G.K. Interleukin involvement in anterior pituitary cell growth regulation: effects of IL-2 and IL-6. Endocrinology 132: 459, 1993.PubMedGoogle Scholar
  21. 21.
    Inoue K., Hattori M., Sakai T., Inukai S., Fujimoto N., Ito A. Establishment of a series of pituitary clonal cell lines differing in morphology, hormones secretion, and response to estrogen. Endocrinology 126: 2313, 1990.PubMedCrossRefGoogle Scholar
  22. 22.
    Bordner M., Karin M. A pituitary specific trans-acting can stimulate transcription from the growth hormone promoter in extracts of nonexpressing cells. Cell 50: 267, 1987.CrossRefGoogle Scholar
  23. 23.
    Matsuda T., Hirano T., Kishimoto T. Establishment of an interleukin-6 (IL-6)/B cell stimulatory factor 2 dependent cell line and preparation of anti-IL-6 monoclonal antibodies. Eur. J. Immunol. 78: 951, 1988.CrossRefGoogle Scholar
  24. 24.
    Bolton A.E., Hunter W.M. The labelling of proteins to high specific radioactivities by conjugation to a 125|-containing acylating agent. Biochem. J. 133: 529, 1973.PubMedCentralPubMedGoogle Scholar
  25. 25.
    Taga T., Kawanishi Y., Hardy R.R., Hirano T., Kishimoto T. Receptors for B cell stimulatory factor 2. J. Exp. Med. 166: 967, 1987.PubMedCrossRefGoogle Scholar
  26. 26.
    Scatchard G. The attractions of proteins for small molecules and ions. Ann. NY Acad. Sci. 51: 660, 1949.CrossRefGoogle Scholar
  27. 27.
    Calvo J.C., Padicella J.P., Charreau E.H. Measurement of specific radioactivities in labelled hormones by self-displacement analysis. Biochem. J. 212: 259, 1983.PubMedCentralPubMedGoogle Scholar
  28. 28.
    Yamaguchi M., Sakata M., Matsuzaki N., Koike K., Miyake A., Tanizawa O. Induction by tumor necrosis factor-alpha of rapid release of immunoreactive and bioactive luteinizing hormone from rat pituitary cells in vitro. Neuroendocrinology 52: 468, 1990.PubMedCrossRefGoogle Scholar
  29. 29.
    Levy Y., Tsapis A., Brouet J.C. Interleukin-6 antisense oligonucleotides inhibit the growth of human myeloma cell lines. J. Clin. Invest. 88: 696, 1991.PubMedCentralPubMedCrossRefGoogle Scholar
  30. 30.
    Miki S., Iwano M., Miki Y. Interleukin-6 (IL-6) functions as an in vitro autocrine growth factor in renal cell carcinoma. FEBS Lett. 250: 607, 1989.PubMedCrossRefGoogle Scholar
  31. 31.
    Ferrare N., Schweigerer L., Neufelo G., Mitchell R., Gospodarowicz D. Pituitary follicular cells produce basic fibroblast growth factor. Proc. Natl. Acad. Sci. USA 84: 5773, 1987.CrossRefGoogle Scholar
  32. 32.
    Gospodarowicz D., Lau K. Pituitary follicular cells secrete both vascular endothelial growth factor and follistatin. Biochem. Biophys. Res. Commun. 165: 292, 1987.CrossRefGoogle Scholar
  33. 33.
    Baes M., Allaerts W., Denef C. Evidence of functional communication between folliculo-stellate cells and hormone-secreting cells in perifused anterior pituitary cell aggregates. Endocrinology 120: 685, 1987.PubMedCrossRefGoogle Scholar
  34. 34.
    Ishikawa H., Nogami H., Shirasawa N. Novel clonal strains from adult rat anterior pituitary producing S-100 protein. Nature 303: 711, 1983.PubMedCrossRefGoogle Scholar
  35. 35.
    Schechter J., Ahmad N., Weiner R. Activation of anterior pituitary folliculo-stellate cells in the formation of estrogen-induced prolactin-secreting tumors. Neuroendocrinology 48: 569, 1988.PubMedCrossRefGoogle Scholar
  36. 36.
    Hofler H., Walter G.F., Denk H. Immunohistochemistry of folliculo-stellate cells in normal human adenohypophyses and in pituitary adenomas. Acta Neuropathol. (Berl.) 65: 35, 1984.CrossRefGoogle Scholar
  37. 37.
    Iwaki T., Kondo A., Takeshita I., Nakagaki H., Kitamura K., Tateishi J. Proliferating potential of folliculo-stellate cells in human pituitary adenomas. Acta Neuropathol. (Berl.) 71: 233, 1986.CrossRefGoogle Scholar
  38. 38.
    Velkeniers B., Vergani P., Trouillas J., D’Haens J., Hooghe R.J., Hooghe-Peters E.L. Expression of IL-6 mRNA in normal rat and human pituitaries and in human pituitary adenomas. J. Histochem. Cytochem. 42: 67, 1994.PubMedCrossRefGoogle Scholar
  39. 39.
    Hama T., Kushima Y., Miyamoto M., Kubota M., Takei N., Hatanaka H. Interleukin-6 improves the survival of mesencephalic catecholaminergic and septal cholinergic neurons from postnatal, two-week-old rats in cultures. Neuroscience 40: 445, 1991.PubMedCrossRefGoogle Scholar
  40. 40.
    Hama T., Miyamoto M., Tsukui H., Nishio C., Hatanaka H. Interleukin-6 as a neurotrophic factor for promoting the survival of cultured basal forebrain cholinergic neurons from postnatal rats. Neurosci. Lett. 104: 340, 1989.PubMedCrossRefGoogle Scholar
  41. 41.
    Ohmichi M., Hirota K., Koike K., Kurachi H., Ohtsuka S., Matsuzaki N., Yamaguchi M., Miyake A., Tanizawa O. Binding sites for interleukin-6 in the anterior pituitary gland. Neuroendocrinology 55: 199, 1992.PubMedCrossRefGoogle Scholar
  42. 42.
    Taga T., Hibi M., Hirata Y., Yamasaki K., Yasukawa K., Matsuda T., Hirano T., Kishimoto T. Interleukin-6 triggers the association of its receptor with a possible signal transducer, gp 130. Cell 58: 573, 1989.PubMedCrossRefGoogle Scholar
  43. 43.
    Hibi M., Murakami M., Saito M., Hirano T., Taga T., Kishimoto T. Molecular cloning and expression of an IL-6 signal tranducer, gp130. Cell 63: 1149, 1990.PubMedCrossRefGoogle Scholar
  44. 44.
    Fukunaga R., Ikeda E., Pan C., Seto Y., Nagata S. Functional domains of the granulocyte colony-stimulating factor receptor. EMBO J. 70: 2855, 1991.Google Scholar

Copyright information

© Italian Society of Endocrinology (SIE) 1995

Authors and Affiliations

  • T. Sawada
    • 1
  • Koji Koike
    • 1
  • Y. Kanda
    • 1
  • H. Ikegami
    • 1
  • H. Jikihara
    • 1
  • T. Maeda
    • 1
  • Y. Osako
    • 1
  • K. Hirota
    • 1
  • A. Miyake
    • 1
  1. 1.Department of Obstetrics and GynecologyOsaka University Medical SchoolSuita, OsakaJapan

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