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. 1996 Feb 20;93(4):1406–1411. doi: 10.1073/pnas.93.4.1406

Correlation between the effects of bombesin antagonists on cell proliferation and intracellular calcium concentration in Swiss 3T3 and HT-29 cell lines.

F F Casanueva 1, F R Perez 1, X Casabiell 1, J P Camiña 1, R Z Cai 1, A V Schally 1
PMCID: PMC39951  PMID: 8643644

Abstract

Bombesin (BN) acts as an autocrine mitogen in various human cancers. Several pseudononapeptide BN-(6-14) analogs with a reduced peptide bond between positions 13 and 14 have been shown to suppress the mitogenic activity of BN or gastrin-releasing peptide (GRP) when assessed by radioreceptor or proliferation assays and may have significant clinical applications. The search for potent and safe BN antagonists requires the evaluation of a large series of analogs in radioreceptor and proliferation assays. In this paper, we report that the ability of BN analogs to inhibit BN-induced calcium transients in Swiss 3T3 cells shows a high correlation with their inhibitory potency as evaluated by classical proliferation tests. The assay of calcium transients allows a rapid characterization of new BN analogs (in terms of minutes rather than days) and can be adapted as a labor and cost-effective screening step in the selection of potentially relevant BN antagonists for further characterization in cell proliferation systems. We also observed that results from the assay of calcium transients in Swiss 3T3 cells can be correlated with the results of the proliferative response in HT-29 cells, a cell line that does not seem to use the same early transmembrane ionic signal system. This result suggests that the calcium pathway is not mandatory for triggering cell division by the BN receptor.

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Selected References

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  1. Berridge M. J., Irvine R. F. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. doi: 10.1038/341197a0. [DOI] [PubMed] [Google Scholar]
  2. Cai R. Z., Radulovic S., Pinski J., Nagy A., Redding T. W., Olsen D. B., Schally A. V. Pseudononapeptide bombesin antagonists containing C-terminal Trp or Tpi. Peptides. 1992 Mar-Apr;13(2):267–271. doi: 10.1016/0196-9781(92)90107-e. [DOI] [PubMed] [Google Scholar]
  3. Cai R. Z., Reile H., Armatis P., Schally A. V. Potent bombesin antagonists with C-terminal Leu-psi(CH2-N)-Tac-NH2 or its derivatives. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12664–12668. doi: 10.1073/pnas.91.26.12664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carney D. N., Cuttitta F., Moody T. W., Minna J. D. Selective stimulation of small cell lung cancer clonal growth by bombesin and gastrin-releasing peptide. Cancer Res. 1987 Feb 1;47(3):821–825. [PubMed] [Google Scholar]
  5. Casabiell X., Pandiella A., Casanueva F. F. Regulation of epidermal-growth-factor-receptor signal transduction by cis-unsaturated fatty acids. Evidence for a protein kinase C-independent mechanism. Biochem J. 1991 Sep 15;278(Pt 3):679–687. doi: 10.1042/bj2780679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Casabiell X., Zugaza J. L., Pombo C. M., Pandiella A., Casanueva F. F. Oleic acid blocks epidermal growth factor-activated early intracellular signals without altering the ensuing mitogenic response. Exp Cell Res. 1993 Apr;205(2):365–373. doi: 10.1006/excr.1993.1099. [DOI] [PubMed] [Google Scholar]
  7. Coy D. H., Heinz-Erian P., Jiang N. Y., Sasaki Y., Taylor J., Moreau J. P., Wolfrey W. T., Gardner J. D., Jensen R. T. Probing peptide backbone function in bombesin. A reduced peptide bond analogue with potent and specific receptor antagonist activity. J Biol Chem. 1988 Apr 15;263(11):5056–5060. [PubMed] [Google Scholar]
  8. Coy D. H., Taylor J. E., Jiang N. Y., Kim S. H., Wang L. H., Huang S. C., Moreau J. P., Gardner J. D., Jensen R. T. Short-chain pseudopeptide bombesin receptor antagonists with enhanced binding affinities for pancreatic acinar and Swiss 3T3 cells display strong antimitotic activity. J Biol Chem. 1989 Sep 5;264(25):14691–14697. [PubMed] [Google Scholar]
  9. Cuttitta F., Carney D. N., Mulshine J., Moody T. W., Fedorko J., Fischler A., Minna J. D. Bombesin-like peptides can function as autocrine growth factors in human small-cell lung cancer. 1985 Aug 29-Sep 4Nature. 316(6031):823–826. doi: 10.1038/316823a0. [DOI] [PubMed] [Google Scholar]
  10. Erusalimsky J. D., Friedberg I., Rozengurt E. Bombesin, diacylglycerols, and phorbol esters rapidly stimulate the phosphorylation of an Mr = 80,000 protein kinase C substrate in permeabilized 3T3 cells. Effect of guanine nucleotides. J Biol Chem. 1988 Dec 15;263(35):19188–19194. [PubMed] [Google Scholar]
  11. Gibbs J., Fauser D. J., Rowe E. A., Rolls B. J., Rolls E. T., Maddison S. P. Bombesin suppresses feeding in rats. Nature. 1979 Nov 8;282(5735):208–210. doi: 10.1038/282208a0. [DOI] [PubMed] [Google Scholar]
  12. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  13. Hirai M., Ishizuka J., Hirai A., Bold R. J., Townsend C. M., Jr, Thompson J. C. Bombesin stimulates intracellular Ca2+ mobilization but not proliferation on human colon cancer cells. Life Sci. 1993;53(24):1859–1865. doi: 10.1016/0024-3205(93)90493-m. [DOI] [PubMed] [Google Scholar]
  14. Lopez-Rivas A., Adelberg E. A., Rozengurt E. Intracellular K+ and the mitogenic response of 3T3 cells to peptide factors in serum-free medium. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6275–6279. doi: 10.1073/pnas.79.20.6275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mendoza S. A., Schneider J. A., Lopez-Rivas A., Sinnett-Smith J. W., Rozengurt E. Early events elicited by bombesin and structurally related peptides in quiescent Swiss 3T3 cells. II. Changes in Na+ and Ca2+ fluxes, Na+/K+ pump activity, and intracellular pH. J Cell Biol. 1986 Jun;102(6):2223–2233. doi: 10.1083/jcb.102.6.2223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Michell R. H. Inositol lipids and phosphates. Curr Opin Cell Biol. 1989 Apr;1(2):201–205. doi: 10.1016/0955-0674(89)90087-2. [DOI] [PubMed] [Google Scholar]
  17. Milovanovic S. R., Radulovic S., Groot K., Schally A. V. Inhibition of growth of PC-82 human prostate cancer line xenografts in nude mice by bombesin antagonist RC-3095 or combination of agonist [D-Trp6]-luteinizing hormone-releasing hormone and somatostatin analog RC-160. Prostate. 1992;20(4):269–280. doi: 10.1002/pros.2990200403. [DOI] [PubMed] [Google Scholar]
  18. Mohammadi M., Dionne C. A., Li W., Li N., Spivak T., Honegger A. M., Jaye M., Schlessinger J. Point mutation in FGF receptor eliminates phosphatidylinositol hydrolysis without affecting mitogenesis. Nature. 1992 Aug 20;358(6388):681–684. doi: 10.1038/358681a0. [DOI] [PubMed] [Google Scholar]
  19. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  20. Qin Y., Halmos G., Cai R. Z., Szoke B., Ertl T., Schally A. V. 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. 1994;120(9):519–528. doi: 10.1007/BF01221028. [DOI] [PubMed] [Google Scholar]
  21. Radulovic S., Cai R. Z., Serfozo P., Groot K., Redding T. W., Pinski J., Schally A. V. 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. 1991 Dec;38(6):593–600. doi: 10.1111/j.1399-3011.1991.tb01545.x. [DOI] [PubMed] [Google Scholar]
  22. Radulovic S., Miller G., Schally A. V. Inhibition of growth of HT-29 human colon cancer xenografts in nude mice by treatment with bombesin/gastrin releasing peptide antagonist (RC-3095). Cancer Res. 1991 Nov 1;51(21):6006–6009. [PubMed] [Google Scholar]
  23. Rozengurt E., Sinnett-Smith J. Bombesin stimulation of DNA synthesis and cell division in cultures of Swiss 3T3 cells. Proc Natl Acad Sci U S A. 1983 May;80(10):2936–2940. doi: 10.1073/pnas.80.10.2936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Seedorf K., Millauer B., Kostka G., Schlessinger J., Ullrich A. Differential effects of carboxy-terminal sequence deletions on platelet-derived growth factor receptor signaling activities and interactions with cellular substrates. Mol Cell Biol. 1992 Oct;12(10):4347–4356. doi: 10.1128/mcb.12.10.4347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shirahige Y., Cai R. Z., Szepeshazi K., Halmos G., Pinski J., Groot K., Schally A. V. Inhibitory effect of bombesin/gastrin-releasing peptide (GRP) antagonists RC-3950-II and RC-3095 on MCF-7 MIII human breast cancer xenografts in nude mice. Biomed Pharmacother. 1994;48(10):465–472. doi: 10.1016/0753-3322(94)90007-8. [DOI] [PubMed] [Google Scholar]
  26. Swope S. L., Schonbrunn A. Bombesin stimulates insulin secretion by a pancreatic islet cell line. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1822–1826. doi: 10.1073/pnas.81.6.1822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Szepeshazi K., Schally A. V., Cai R. Z., Radulovic S., Milovanovic S., Szoke B. Inhibitory effect of bombesin/gastrin-releasing peptide antagonist RC-3095 and high dose of somatostatin analogue RC-160 on nitrosamine-induced pancreatic cancers in hamsters. Cancer Res. 1991 Nov 1;51(21):5980–5986. [PubMed] [Google Scholar]
  28. Szepeshazi K., Schally A. V., Groot K., Halmos G. Effect of bombesin, gastrin-releasing peptide (GRP)(14-27) and bombesin/GRP receptor antagonist RC-3095 on growth of nitrosamine-induced pancreatic cancers in hamsters. Int J Cancer. 1993 May 8;54(2):282–289. doi: 10.1002/ijc.2910540220. [DOI] [PubMed] [Google Scholar]
  29. Szepeshazi K., Schally A. V., Halmos G., Groot K., Radulovic S. 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. 1992 Dec 16;84(24):1915–1922. doi: 10.1093/jnci/84.24.1915. [DOI] [PubMed] [Google Scholar]
  30. Taché Y., Vale W., Rivier J., Brown M. Brain regulation of gastric secretion: influence of neuropeptides. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5515–5519. doi: 10.1073/pnas.77.9.5515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Takuwa N., Takuwa Y., Bollag W. E., Rasmussen H. The effects of bombesin on polyphosphoinositide and calcium metabolism in Swiss 3T3 cells. J Biol Chem. 1987 Jan 5;262(1):182–188. [PubMed] [Google Scholar]
  32. Wang L. H., Coy D. H., Taylor J. E., Jiang N. Y., Kim S. H., Moreau J. P., Huang S. C., Mantey S. A., Frucht H., Jensen R. T. Desmethionine alkylamide bombesin analogues: a new class of bombesin receptor antagonists with potent antisecretory activity in pancreatic acini and antimitotic activity in Swiss 3T3 cells. Biochemistry. 1990 Jan 23;29(3):616–622. doi: 10.1021/bi00455a004. [DOI] [PubMed] [Google Scholar]
  33. Willey J. C., Lechner J. F., Harris C. C. Bombesin and the C-terminal tetradecapeptide of gastrin-releasing peptide are growth factors for normal human bronchial epithelial cells. Exp Cell Res. 1984 Jul;153(1):245–248. doi: 10.1016/0014-4827(84)90466-x. [DOI] [PubMed] [Google Scholar]
  34. Woll P. J., Rozengurt E. [D-Arg1,D-Phe5,D-Trp7,9,Leu11]substance P, a potent bombesin antagonist in murine Swiss 3T3 cells, inhibits the growth of human small cell lung cancer cells in vitro. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1859–1863. doi: 10.1073/pnas.85.6.1859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Yano T., Pinski J., Szepeshazi K., Halmos G., Radulovic S., Groot K., Schally A. V. Inhibitory effect of bombesin/gastrin-releasing peptide 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. 1994 Feb 15;73(4):1229–1238. doi: 10.1002/1097-0142(19940215)73:4<1229::aid-cncr2820730417>3.0.co;2-1. [DOI] [PubMed] [Google Scholar]
  37. Zugaza J. L., Casabiell X. A., Bokser L., Casanueva F. F. Oleic acid blocks EGF-induced [Ca2+]i release without altering cellular metabolism in fibroblast EGFR T17. Biochem Biophys Res Commun. 1995 Feb 6;207(1):105–110. doi: 10.1006/bbrc.1995.1159. [DOI] [PubMed] [Google Scholar]
  38. Zugaza J. L., Casabiell X. A., Bokser L., Eiras A., Beiras A., Casanueva F. F. Pretreatment with oleic acid accelerates the entrance into the mitotic cycle of EGF-stimulated fibroblasts. Exp Cell Res. 1995 Jul;219(1):54–63. doi: 10.1006/excr.1995.1204. [DOI] [PubMed] [Google Scholar]

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