Skip to main content
NCBI 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 1;91(5):1701–1705. doi: 10.1073/pnas.91.5.1701

Inhibition of human epithelial ovarian cancer cell growth in vitro by agonistic and antagonistic analogues of luteinizing hormone-releasing hormone.

T Yano 1, J Pinski 1, S Radulovic 1, A V Schally 1
PMCID: PMC43231  PMID: 8127868

Abstract

In this study, we investigated the effects of luteinizing hormone-releasing hormone (LH-RH) agonist [D-Trp6]LH-RH, LH-RH antagonist [Ac-D-Nal(2)1,D-Phe(pCl)2,D-Pal(3)3,D-Cit6,D-Ala10]LH-RH (SB-75), and estradiol on the growth of human epithelial ovarian cancer cell line OV-1063. Cells were cultured under estrogen-deprived conditions. Estradiol inhibited cell proliferation, as measured by cell number at 10(-9)-10(-7) M and [3H]thymidine incorporation into DNA at 10(-13)-10(-8) M. Both LH-RH analogs inhibited cell growth dose dependently in the range 10(-8)-10(-5) M, but SB-75 induced a greater growth inhibition than [D-Trp6]LH-RH. In OV-1063 cells, 125I-labeled [D-Trp6]LH-RH was bound to one class of specific, saturable binding sites with high affinity (Kd = 1.4 +/- 0.3 nM) and low capacity (4000 binding sites per cell). 125I-labeled [D-Trp6]LH-RH could be displaced by unlabeled [D-Trp6]LH-RH and SB-75, suggesting that both analogs are bound to the same receptor on OV-1063 cells. Ligand binding was dependent on time and temperature. Receptor internalization assay showed that the ligand-receptor complex was internalized at 37 degrees C, which indicates the presence of biologically active LH-RH receptors on OV-1063 cells. These results suggest that estradiol and LH-RH analogs can suppress the growth of OV-1063 human epithelial ovarian cancer cells by a direct action and that the inhibitory effect of LH-RH analogs is mediated through the high-affinity LH-RH receptors.

Full text

PDF
1701

Selected References

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

  1. Bajusz S., Csernus V. J., Janaky T., Bokser L., Fekete M., Schally A. V. New antagonists of LHRH. II. Inhibition and potentiation of LHRH by closely related analogues. Int J Pept Protein Res. 1988 Dec;32(6):425–435. doi: 10.1111/j.1399-3011.1988.tb01373.x. [DOI] [PubMed] [Google Scholar]
  2. Berthois Y., Katzenellenbogen J. A., Katzenellenbogen B. S. Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2496–2500. doi: 10.1073/pnas.83.8.2496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blankenstein M. A., Henkelman M. S., Klijn J. G. Direct inhibitory effect of a luteinizing hormone-releasing hormone agonist on MCF-7 human breast cancer cells. Eur J Cancer Clin Oncol. 1985 Dec;21(12):1493–1499. doi: 10.1016/0277-5379(85)90244-5. [DOI] [PubMed] [Google Scholar]
  4. Boring C. C., Squires T. S., Tong T. Cancer statistics, 1992. CA Cancer J Clin. 1992 Jan-Feb;42(1):19–38. doi: 10.3322/canjclin.42.1.19. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Bruckner H. W., Motwani B. T. Treatment of advanced refractory ovarian carcinoma with a gonadotropin-releasing hormone analogue. Am J Obstet Gynecol. 1989 Nov;161(5):1216–1218. doi: 10.1016/0002-9378(89)90669-8. [DOI] [PubMed] [Google Scholar]
  7. Csernus V. J., Szende B., Groot K., Redding T. W., Schally A. V. Development of radioimmunoassay for a potent luteinizing hormone-releasing hormone antagonist. Evaluation of serum levels after injection of [Ac-3-(2-naphthyl)-D-Ala1, D-Phe(pCl)2, 3-(3-pyridyl)-D-Ala3, D-Cit6, D-Ala10] LHRH. Arzneimittelforschung. 1990 Feb;40(2 Pt 1):111–118. [PubMed] [Google Scholar]
  8. Eidne K. A., Flanagan C. A., Harris N. S., Millar R. P. Gonadotropin-releasing hormone (GnRH)-binding sites in human breast cancer cell lines and inhibitory effects of GnRH antagonists. J Clin Endocrinol Metab. 1987 Mar;64(3):425–432. doi: 10.1210/jcem-64-3-425. [DOI] [PubMed] [Google Scholar]
  9. Emons G., Pahwa G. S., Brack C., Sturm R., Oberheuser F., Knuppen R. Gonadotropin releasing hormone binding sites in human epithelial ovarian carcinomata. Eur J Cancer Clin Oncol. 1989 Feb;25(2):215–221. doi: 10.1016/0277-5379(89)90011-4. [DOI] [PubMed] [Google Scholar]
  10. Fekete M., Bajusz S., Groot K., Csernus V. J., Schally A. V. Comparison of different agonists and antagonists of luteinizing hormone-releasing hormone for receptor-binding ability to rat pituitary and human breast cancer membranes. Endocrinology. 1989 Feb;124(2):946–955. doi: 10.1210/endo-124-2-946. [DOI] [PubMed] [Google Scholar]
  11. Haigler H. T. Receptor-mediated endocytosis of epidermal growth factor. Methods Enzymol. 1983;98:283–290. doi: 10.1016/0076-6879(83)98155-7. [DOI] [PubMed] [Google Scholar]
  12. Hazum E., Cuatrecasas P., Marian J., Conn P. M. Receptor-mediated internalization of fluorescent gonadotropin-releasing hormone by pituitary gonadotropes. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6692–6695. doi: 10.1073/pnas.77.11.6692. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hershkovitz E., Marbach M., Bosin E., Levy J., Roberts C. T., Jr, LeRoith D., Schally A. V., Sharoni Y. Luteinizing hormone-releasing hormone antagonists interfere with autocrine and paracrine growth stimulation of MCF-7 mammary cancer cells by insulin-like growth factors. J Clin Endocrinol Metab. 1993 Oct;77(4):963–968. doi: 10.1210/jcem.77.4.8408472. [DOI] [PubMed] [Google Scholar]
  14. Horowitz A. T., Treves A. J., Voss R., Okon E., Fuks Z., Davidson L., Biran S. A new human ovarian carcinoma cell line: establishment and analysis of tumor-associated markers. Oncology. 1985;42(5):332–337. doi: 10.1159/000226056. [DOI] [PubMed] [Google Scholar]
  15. Jäger W., Wildt L., Lang N. Some observations on the effect of a GnRH analog in ovarian cancer. Eur J Obstet Gynecol Reprod Biol. 1989 Aug;32(2):137–148. doi: 10.1016/0028-2243(89)90195-0. [DOI] [PubMed] [Google Scholar]
  16. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  17. Nakano R., Kitayama S., Yamoto M., Shima K., Ooshima A. Localization of gonadotropin binding sites in human ovarian neoplasms. Am J Obstet Gynecol. 1989 Oct;161(4):905–910. doi: 10.1016/0002-9378(89)90749-7. [DOI] [PubMed] [Google Scholar]
  18. Nash J. D., Ozols R. F., Smyth J. F., Hamilton T. C. Estrogen and anti-estrogen effects on the growth of human epithelial ovarian cancer in vitro. Obstet Gynecol. 1989 Jun;73(6):1009–1016. doi: 10.1097/00006250-198906000-00021. [DOI] [PubMed] [Google Scholar]
  19. Pahwa G. S., Vollmer G., Knuppen R., Emons G. Photoaffinity labelling of gonadotropin releasing hormone binding sites in human epithelial ovarian carcinomata. Biochem Biophys Res Commun. 1989 Jun 30;161(3):1086–1092. doi: 10.1016/0006-291x(89)91354-5. [DOI] [PubMed] [Google Scholar]
  20. Parmar H., Nicoll J., Stockdale A., Cassoni A., Phillips R. H., Lightman S. L., Schally A. V. Advanced ovarian carcinoma: response to the agonist D-Trp-6-LHRH. Cancer Treat Rep. 1985 Nov;69(11):1341–1342. [PubMed] [Google Scholar]
  21. Parmar H., Rustin G., Lightman S. L., Phillips R. H., Hanham I. W., Schally A. V. Response to D-Trp-6-luteinising hormone releasing hormone (Decapeptyl) microcapsules in advanced ovarian cancer. Br Med J (Clin Res Ed) 1988 Apr 30;296(6631):1229–1229. doi: 10.1136/bmj.296.6631.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rao B. R., Slotman B. J. Endocrine factors in common epithelial ovarian cancer. Endocr Rev. 1991 Feb;12(1):14–26. doi: 10.1210/edrv-12-1-14. [DOI] [PubMed] [Google Scholar]
  23. Segal-Abramson T., Kitroser H., Levy J., Schally A. V., Sharoni Y. Direct effects of luteinizing hormone-releasing hormone agonists and antagonists on MCF-7 mammary cancer cells. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2336–2339. doi: 10.1073/pnas.89.6.2336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sharoni Y., Bosin E., Miinster A., Levy J., Schally A. V. Inhibition of growth of human mammary tumor cells by potent antagonists of luteinizing hormone-releasing hormone. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1648–1651. doi: 10.1073/pnas.86.5.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Simon W. E., Albrecht M., Hänsel M., Dietel M., Hölzel F. Cell lines derived from human ovarian carcinomas: growth stimulation by gonadotropic and steroid hormones. J Natl Cancer Inst. 1983 May;70(5):839–845. [PubMed] [Google Scholar]
  26. Thompson M. A., Adelson M. D., Kaufman L. M. Lupron retards proliferation of ovarian epithelial tumor cells cultured in serum-free medium. J Clin Endocrinol Metab. 1991 May;72(5):1036–1041. doi: 10.1210/jcem-72-5-1036. [DOI] [PubMed] [Google Scholar]
  27. Wimalasena J., Meehan D., Cavallo C. Human epithelial ovarian cancer cell steroid secretion and its control by gonadotropins. Gynecol Oncol. 1991 Apr;41(1):56–63. doi: 10.1016/0090-8258(91)90255-4. [DOI] [PubMed] [Google Scholar]
  28. Yano T., Korkut E., Pinski J., Szepeshazi K., Milovanovic S., Groot K., Clarke R., Comaru-Schally A. M., Schally A. V. Inhibition of growth of MCF-7 MIII human breast carcinoma in nude mice by treatment with agonists or antagonists of LH-RH. Breast Cancer Res Treat. 1992;21(1):35–45. doi: 10.1007/BF01811962. [DOI] [PubMed] [Google Scholar]
  29. Yano T., Pinski J., Groot K., Schally A. V. Stimulation by bombesin and inhibition by bombesin/gastrin-releasing peptide antagonist RC-3095 of growth of human breast cancer cell lines. Cancer Res. 1992 Aug 15;52(16):4545–4547. [PubMed] [Google Scholar]
  30. Yano T., Pinski J., Szepeshazi K., Milovanovic S. R., Groot K., Schally A. V. Effect of microcapsules of luteinizing hormone-releasing hormone antagonist SB-75 and somatostatin analog RC-160 on endocrine status and tumor growth in the Dunning R-3327H rat prostate cancer model. Prostate. 1992;20(4):297–310. doi: 10.1002/pros.2990200405. [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