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

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Abstract

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.

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References

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  14. Hirano T. Interleukin 6 (IL-6) and its receptor: Their role in plasma cell neoplasias. Int. J. Cell. Cloning 9: 166, 1991.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  Google Scholar 

  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.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  Google Scholar 

  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.

    Article  Google Scholar 

  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.

    PubMed Central  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  26. Scatchard G. The attractions of proteins for small molecules and ions. Ann. NY Acad. Sci. 51: 660, 1949.

    Article  CAS  Google Scholar 

  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.

    PubMed Central  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  Google Scholar 

  32. Gospodarowicz D., Lau K. Pituitary follicular cells secrete both vascular endothelial growth factor and follistatin. Biochem. Biophys. Res. Commun. 165: 292, 1987.

    Article  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  34. Ishikawa H., Nogami H., Shirasawa N. Novel clonal strains from adult rat anterior pituitary producing S-100 protein. Nature 303: 711, 1983.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  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.

    Article  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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.

    Article  PubMed  CAS  Google Scholar 

  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 

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Osaka University Medical School, 2-2, Yamadaoka, Suita, Osaka, 565, Japan

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

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  2. Koji Koike
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Sawada, T., Koike, K., Kanda, Y. et al. Interleukin-6 stimulates cell proliferation of rat pituitary clonal cell lines in vitro . J Endocrinol Invest 18, 83–90 (1995). https://doi.org/10.1007/BF03349706

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