Regular Article

British Journal of Cancer (2000) 83, 906–913. doi:10.1054/bjoc.2000.1374 www.bjcancer.com
Published online 5 September 2000

Inhibition of growth of OV-1063 human epithelial ovarian cancers and c-jun and c-fos oncogene expression by bombesin antagonists

I Chatzistamou1,2, A V Schally1,2, B Sun1,2, P Armatis1 and K Szepeshazi1,2

  1. 1Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center and Section of Experimental Medicine, Tulane University School of Medicine, New Orleans, LA 70112, USA
  2. 2Department of Medicine, Tulane University School of Medicine, New Orleans, LA 70112, USA

Received 29 November 1999; Revised 17 May 2000; Accepted 23 May 2000.

Top

Abstract

Receptors for bombesin are present on human ovarian cancers and bombesin-like peptides could function as growth factors in this carcinoma. Therefore, we investigated the effects of bombesin/gastrin-releasing peptide (GRP) antagonists RC-3940-II and RC-3095 on the growth of human ovarian carcinoma cell line OV-1063, xenografted into nude mice. Treatment with RC-3940-II at doses of 10 mug and 20 mug per day s.c. decreased tumour volume by 60.9% (P< 0.05) and 73.5% (P< 0.05) respectively, after 25 days, compared to controls. RC-3095 at a dose of 20 mug per day reduced the volume of OV-1063 tumours by 47.7% (P = 0.15). In comparison, luteinizing hormone-releasing hormone (LH-RH) antagonist Cetrorelix at a dose of 100 mug per day caused a 64.2% inhibition (P< 0.05). RT-PCR analysis showed that OV-1063 tumours expressed mRNA for bombesin receptor subtypes BRS-1, BRS-2, and BRS-3. In OV-1063 cells cultured in vitro, GRP(14–27) induced the expression of mRNA for c-jun and c-fos oncogenes in a time-dependent manner. Antagonist RC-3940-II inhibited the stimulatory effect of GRP(14–27) on c-jun and c-fos in vitro. In vivo, the levels of c-jun and c-fos mRNA in OV-1063 tumours were decreased by 43% (P< 0.05) and 45% (P = 0.05) respectively, after treatment with RC-3940-II at 20 mug per day. Exposure of OV-1063, UCI-107 and ES-2 ovarian carcinoma cells to RC-3940-II at 1 muM concentration for 24 h in vitro, extended the latency period for the development of palpable tumours in nude mice. Our results indicate that antagonists of bombesin/GRP inhibit the growth of OV-1063 ovarian cancers by mechanisms that probably involve the downregulation of c-jun and c-fos proto-oncogenes. © 2000 Cancer Research Campaign

Keywords:

cancer therapy, bombesin/GRP antagonists, LH-RH antagonist, ovarian tumours, c-jun, c-fos

Top

References

  1. Bajusz S, Csernus VJ, Janaky T, Bokser L, Fekete M and Schally AV (1988) New antagonists of LH-RH: II. Inhibition and potentiation of LH-RH by closely related analogues. Int J Peptide Protein Res 32: 425–435
  2. Bernhardt G, Reile H, Birnbock H, Sprub T and Schonenberger H (1992) Standardized kinetic microassay to quantify differential chemosensitivity on the basis of proliferative activity. J Cancer Res Clin Oncol 118: 35–43 | PubMed | ISI | ChemPort |
  3. Bologna MC, Festuccia C, Muzi P, Biordi L and Ciomei M (1989) Bombesin stimulates growth of human prostatic cancer cells in vitro. Cancer 63: 1714–1720 | Article | PubMed | ISI | ChemPort |
  4. Cai RZ, Reile H, Armatis P and Schally AV (1994) Potent bombesin antagonists with C-terminal Leupsi(CH2N)Tac-NH2or its derivatives. Proc Natl Acad Sci USA 91: 12664–12668 | Article | PubMed | ChemPort |
  5. Carney DN, Cuttitta F, Moody TW and Minna JD (1987) Selective stimulation of small cell lung cancer clonal growth by bombesin and gastrin-releasing peptides. Cancer Res 47: 821–825 | PubMed |
  6. Chegini N, Rong H, Dou Q, Kipersztok C and Williams ARS (1996) Gonadotropin-releasing hormone (GnRH) and GnRH receptor gene expression in human myometrium and leiomyomata and the direct action of GNRH analogs on myometrial smooth muscle cells and interaction with ovarian steroids in vitro. J Clin Endocrinol Metab 81: 3215–3221
  7. Cuttitta F, Carney DN, Mulshine J, Moody TW, Fedorko J, Fishler A and Minna JD (1985) Bombesin-like peptides can function as autocrine growth factors in human small-cell lung cancer. Nature 316: 823–826 | Article | PubMed | ISI | ChemPort |
  8. Draoui M, Chung P, Park M, Birrer M, Jakowlew S and Moody TW (1995) Bombesin stimulates c-fos and c-jun mRNAs in small cell lung cancer cells. Peptides 16: 289–292 | PubMed |
  9. Emons G, Muller V, Ortmann O and Schulz K-D (1998) Effects of LH-RH-analogues on mitogenic signal transduction in cancer cells. J Steroid Biochem Mol Biol 65: 199–206 | Article | PubMed | ChemPort |
  10. Halazonetis TD, Georgopoulos K, Greenberg ME and Leder P (1988) C-jun dimerizes with itself and with c-fos, forming complexes of different DNA binding affinities. Cell 55: 917–924 | Article | PubMed | ISI | ChemPort |
  11. Halmos G, Wittliff JL and Schally AV (1995) Characterization of bombesin/GRP receptors in human breast cancer and their relationship to steroid receptor expression. Cancer Res 53: 280–287
  12. Halmos G and Schally AV (1997) Reduction in receptors for bombesin and epidermal growth factor in xenografts of human small-cell lung cancer after treatment with bombesin antagonist RC-3095. Proc Natl Acad Sci USA 94: 956–960 | PubMed |
  13. Horowitz AT, Treves AJ, Voss R, Okon E, Fuks Z, Davinson L and Biran S (1985) A new human ovarian carcinoma cell line: establishment and analysis of tumour associated markers. Oncology 42: 332–337
  14. Jungwirth A, Galvan G, Pinski J, Halmos G, Szepeshazi K, Cai RZ, Groot K and Schally AV1997 a) Luteinizing hormone-releasing hormone antagonist Cetrorelix (SB-75) and bombesin antagonist RC-3940-II inhibit the growth of androgen-independent PC-3 prostate cancer in nude mice. Prostate 32: 164–172
  15. Jungwirth A, Pinski J, Galvan G, Halmos G, Szepeshazi K, Cai RZ, Groot K, Vadillo-Buenfil M and Schally AV1997 b) Inhibition of growth of androgen-independent DU-145 prostate cancer in vivo by luteinising hormone-releasing hormone antagonist Cetrorelix and bombesin antagonists RC-3940-II and RC-3950-II. Eur J Cancer 33: 1141–1148 | Article | PubMed | ISI | ChemPort |
  16. Jungwirth A, Schally AV, Halmos G, Groot K, Szepeshazi K, Pinski J and Armatis P (1998) Inhibition of the growth of Caki-I human renal adenocarcinoma in vivo by luteinizing hormone-releasing hormone antagonist Cetrorelix, somatostatin analog RC-160, and bombesin antagonist RC-3940-II. Cancer 82: 909–917 | Article | PubMed | ChemPort |
  17. Kiaris H, Schally AV, Sun B, Armatis P and Groot K (1999) Inhibition of growth of human malignant glioblastoma in nude mice by antagonists of bombesin/gastrin-releasing peptide. Oncogene 18: 7168–7173 | Article | PubMed | ChemPort |
  18. Kim HJ, Evers MB, Banker NA, Greeley GH, Hellmich MR, Thompson JC and Townsend CM (1998) Novel expression and regulation of gastrin gene in human ovarian cancer cell line, SW626. Dig Dis Sci 43: 1465–1473
  19. Koppan M, Halmos G, Arencibia JM, Lambarzi N and Schally AV (1998) Bombesin/Gastrin-Releasing Peptide Antagonists RC-3095 and RC-3940-II inhibit tumour growth and decrease the levels and mRNA expression of epidermal growth factor receptors in H-69 small cell lung carcinoma. Cancer 83: 1335–1343 | PubMed | ChemPort |
  20. Landis SH, Murray T, Bolden S and Wingo PA (1999) Cancer Statistics, 1999. CA Cancer J Clin 49: 8–31 | Article | PubMed | ISI | ChemPort |
  21. Levi F, Lucchini F, Negri E, Boyle P and La Vecchia C (1999) Cancer mortality in Europe, 1990–1994, and an overview of trends from 1955–1994. Eur J Cancer 35: 1447–1516
  22. Liebow C, Crean DH, Lee MT, Kamer AR, Mang TS and Schally AV (1994) Synergistic effects of bombesin and epidermal growth factor on cancers. Proc Natl Acad Sci USA 91: 3804–3808 | PubMed | ChemPort |
  23. Markwalder R and Reubi JC (1999) Gastrin-releasing peptide receptors in the human prostate: Relation to neoplastic transformation. Cancer Res 59: 1152–1159 | PubMed | ISI | ChemPort |
  24. Miyazaki M, Lamharzi N, Schally AV, Halmos G, Szepeshazi K, Groot K and Cai RZ (1998) Inhibition of growth of MDA-MB-231 human breast cancer xenografts in nude mice by bombesin/gastrin-releasing peptide (GRP) antagonists RC-3940-II and RC-3095. Eur J Cancer 34: 710–717
  25. Moody TW and Cuttitta F (1993) Growth factor and peptide receptors in small cell lung cancer. Life Sci 52: 1161–1173
  26. Nagalla SR, Barry BJ, Creswick KC, Eden P, Taylor JT and Spindel ER (1995) Cloning of a receptor for amphibian [Phe13]bombesin distinct from the receptor for gastrin-releasing peptide: Identification of a fourth bombesin receptor subtype (BR4). Proc Natl Acad Sci USA 92: 6205–6209 | PubMed | ChemPort |
  27. Qin Y, Ertl T, Cai RZ, Halmos G and Schally AV1994 a) Inhibitory effect of bombesin receptor antagonist RC-3095 on the growth of human pancreatic cancer cells in vivo and in vitro. Cancer Res 54: 1035–1041
  28. Qin Y, Halmos G, Cai RZ, Szoke B, Ertl T and Schally AV1994 b) Bombesin antagonists inhibit in vitro and in vivo growth of human gastric cancer and binding of bombesin to its receptors. J Cancer Res Clin Oncol 120: 519–528
  29. Radulovic S, Cai RZ, Serfoso P, Groot K, Redding TW, Pinski J and Schally AV (1991) Biological effects and receptor binding affinities of new pseudononapeptide bombesin/GRP receptor antagonists with N-terminal D-Trp or D-Tpi. Int J Pept Protein Res 38: 593–600 | PubMed |
  30. Reile H, Cai RZ, Armatis P and Schally AV (1995) New antagonists of bombesin/gastrin-releasing peptide with C-terminal Leupsi(CH2N)Tac-NH2. Int J Oncol 7: 749–754 | ISI | ChemPort |
  31. Schally AV and Comaru-Schally AM (1997) Hypothalamic and other peptide hormones. In Cancer Medicine (4th edn) Holland JF, Frei III E, Bast Jr RC, Kufe DE, Morton DL, Weichselbaum RR (eds), pp 1067–1086. Williams and Wilkins: Baltimore
  32. Shirahige Y, Cai RZ, Szepeshazi K, Halmos G, Pinski J, Groot K and Schally AV (1994) Inhibitory effect of bombesin/gastrin-releasing peptide (GRP) antagonists RC-3950-II and RC-3095 on MCF7-MIII human breast cancer xenografts in nude mice. Biomed Pharmacother 48: 465–472 | Article | PubMed | ChemPort |
  33. Siegfried JM, Han YH, DeMichele MAA, Hunt JD, Gaither AL and Cuttitta F (1994) Production of gastrin-releasin peptide by a non-small cell lung carcinoma cell line adapted to serum-free and growth factor-free conditions. J Biol Chem 269: 8596–8603 | PubMed |
  34. Spindel ER, Giladi E, Segerson TP and Nagalla S (1993) Bombesin-like peptides: of ligands and receptors. Recent Prog Horm Res 48: 365–391
  35. Steel RGD and Torrie J (1976) Principles and Procedures of Statistics. McGraw Hill: New York
  36. Sun B, Schally AV and Halmos G (2000) The presence of receptors for bombesin/ GRP and mRNA for three receptor subtypes in human ovarian epithelial cancers. Reg Peptides 90: 77–84
  37. Sun B, Halmos G, Schally AV, Wang X and Martinez M (2000) The presence of receptors for bombesin/gastrin-releasing peptide and mRNA for 3 receptor subtypes in human prostate cancers. Prostate 42: 295–303
  38. Szepeshazi K, Schally AV, Halmos G, Groot K and Radulovic S (1992) Growth inhibition of estrogen-dependent and estrogen-independent MXT mammary cancers in mice by the bombesin and gastrin-releasing peptide antagonist RC-3095. J Natl Cancer Inst 84: 1915–1922 | PubMed |
  39. Szepeshazi K, Schally AV, Halmos G, Lamharzi N, Groot K and Horvath JE (1997) A single in vivo administration of bombesin antagonist RC-3095 reduces the levels and mRNA expression of epidermal growth factor receptors in MXT mouse mammary cancers. Proc Natl Acad Sci USA 94: 10913–10918 | PubMed |
  40. Tomayko MM and Reynolds CP (1989) Determination of subcutaneous tumour size in athymic (nude) mice. Cancer Chemother Pharmacol 24: 148–154 | Article | PubMed | ISI | ChemPort |
  41. UKCCCR (1998) United Kingdom Coordinating Committee on Cancer Research (UKCCCR) guidelines for the welfare of animals in experimental neoplasia (2nd edn). Br J Cancer 77: 1–10 | PubMed | ISI | ChemPort |
  42. Wang QJ, Knezetic AJ, Schally AV, Pour PM and Adrian T (1996) Bombesin may stimulate proliferation of human pancreatic cancer cells through an autocrine pathway. Int J Cancer 68: 1–7 | PubMed |
  43. Yano T, Pinski J, Groot K and Schally AV (1992) Stimulation by bombesin and inhibition by bombesin/gastrin releasing peptide antagonist RC-3950 of growth of human breast cancer cell lines. Cancer Res 54: 4545–4547
  44. Yano T, Pinski J, Szepeshazi K, Halmos G, Radulovic S, Groot K and Schally AV (1993) Inhibitory effect of bombesin/GRP antagonist RC-3095 and luteinizing hormone-releasing hormone antagonist SB-75 on the growth of MCF-7 MIII human breast cancer xenografts in athymic nude mice. Cancer 73: 1229–1238
  45. Yano T, Pinski J, Halmos G, Szepeshazi K, Groot K and Schally AV (1994) Inhibition of growth of OV-1063 human epithelial ovarian cancer xenografts in nude mice by treatment with luteinizing hormone-releasing hormone antagonist SB-75. Proc Natl Acad Sci USA 91: 7090–7094