Abstract
Bombesins are potent growth factors for murine Swiss 3T3 cells. Using these cells in chemically defined conditions we have been able to characterise the bombesin receptor and the early signals preceding DNA synthesis. We describe two substance P analogues [DArg1, DPro2, DTrp7,9, Leu11] substance P and [DArg1, DPhe5, DTrp7,9, Leu11] substance P which competitively block the binding of bombesins to their receptor and all the events leading to mitogenesis. Bombesins are secreted by human small cell lung cancers (SCLC) and may act as autocrine growth factors for these tumours, so the development of peptide bombesin antagonists could have therapeutic implications. We demonstrate that the antagonists can reversibly inhibit the growth of SCLC in vitro, with relatively little effect on other lung tumours.
Full text
PDF







Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anastasi A., Erspamer V., Bucci M. Isolation and structure of bombesin and alytesin, 2 analogous active peptides from the skin of the European amphibians Bombina and Alytes. Experientia. 1971 Feb 15;27(2):166–167. doi: 10.1007/BF02145873. [DOI] [PubMed] [Google Scholar]
- Bailar J. C., 3rd, Smith E. M. Progress against cancer? N Engl J Med. 1986 May 8;314(19):1226–1232. doi: 10.1056/NEJM198605083141905. [DOI] [PubMed] [Google Scholar]
- Ballester R., Rosen O. M. Fate of immunoprecipitable protein kinase C in GH3 cells treated with phorbol 12-myristate 13-acetate. J Biol Chem. 1985 Dec 5;260(28):15194–15199. [PubMed] [Google Scholar]
- Bergeron J. J., Cruz J., Khan M. N., Posner B. I. Uptake of insulin and other ligands into receptor-rich endocytic components of target cells: the endosomal apparatus. Annu Rev Physiol. 1985;47:383–403. doi: 10.1146/annurev.ph.47.030185.002123. [DOI] [PubMed] [Google Scholar]
- Berridge M. J., Irvine R. F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. doi: 10.1038/312315a0. [DOI] [PubMed] [Google Scholar]
- Bishop R., Martinez R., Weber M. J., Blackshear P. J., Beatty S., Lim R., Herschman H. R. Protein phosphorylation in a tetradecanoyl phorbol acetate-nonproliferative variant of 3T3 cells. Mol Cell Biol. 1985 Sep;5(9):2231–2237. doi: 10.1128/mcb.5.9.2231-2237.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blackshear P. J., Wen L., Glynn B. P., Witters L. A. Protein kinase C-stimulated phosphorylation in vitro of a Mr 80,000 protein phosphorylated in response to phorbol esters and growth factors in intact fibroblasts. Distinction from protein kinase C and prominence in brain. J Biol Chem. 1986 Jan 25;261(3):1459–1469. [PubMed] [Google Scholar]
- Blackshear P. J., Witters L. A., Girard P. R., Kuo J. F., Quamo S. N. Growth factor-stimulated protein phosphorylation in 3T3-L1 cells. Evidence for protein kinase C-dependent and -independent pathways. J Biol Chem. 1985 Oct 25;260(24):13304–13315. [PubMed] [Google Scholar]
- Brown K. D., Blay J., Irvine R. F., Heslop J. P., Berridge M. J. Reduction of epidermal growth factor receptor affinity by heterologous ligands: evidence for a mechanism involving the breakdown of phosphoinositides and the activation of protein kinase C. Biochem Biophys Res Commun. 1984 Aug 30;123(1):377–384. doi: 10.1016/0006-291x(84)90424-8. [DOI] [PubMed] [Google Scholar]
- Carney D. N., Bunn P. A., Jr, Gazdar A. F., Pagan J. A., Minna J. D. Selective growth in serum-free hormone-supplemented medium of tumor cells obtained by biopsy from patients with small cell carcinoma of the lung. Proc Natl Acad Sci U S A. 1981 May;78(5):3185–3189. doi: 10.1073/pnas.78.5.3185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Cirillo D. M., Gaudino G., Naldini L., Comoglio P. M. Receptor for bombesin with associated tyrosine kinase activity. Mol Cell Biol. 1986 Dec;6(12):4641–4649. doi: 10.1128/mcb.6.12.4641-4649.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Corps A. N., Rees L. H., Brown K. D. A peptide that inhibits the mitogenic stimulation of Swiss 3T3 cells by bombesin or vasopressin. Biochem J. 1985 Nov 1;231(3):781–784. doi: 10.1042/bj2310781. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Dicker P., Rozengurt E. Phorbol esters and vasopressin stimulate DNA synthesis by a common mechanism. Nature. 1980 Oct 16;287(5783):607–612. doi: 10.1038/287607a0. [DOI] [PubMed] [Google Scholar]
- Erisman M. D., Linnoila R. I., Hernandez O., DiAugustine R. P., Lazarus L. H. Human lung small-cell carcinoma contains bombesin. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2379–2383. doi: 10.1073/pnas.79.7.2379. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ghatei M. A., Sheppard M. N., Henzen-Logman S., Blank M. A., Polak J. M., Bloom S. R. Bombesin and vasoactive intestinal polypeptide in the developing lung: marked changes in acute respiratory distress syndrome. J Clin Endocrinol Metab. 1983 Dec;57(6):1226–1232. doi: 10.1210/jcem-57-6-1226. [DOI] [PubMed] [Google Scholar]
- Goldstein J. L., Brown M. S., Anderson R. G., Russell D. W., Schneider W. J. Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol. 1985;1:1–39. doi: 10.1146/annurev.cb.01.110185.000245. [DOI] [PubMed] [Google Scholar]
- Hanley M. R., Jackson T. Substance K receptor: return of the magnificent seven. 1987 Oct 29-Nov 4Nature. 329(6142):766–767. doi: 10.1038/329766a0. [DOI] [PubMed] [Google Scholar]
- Heldin C. H., Westermark B. Growth factors: mechanism of action and relation to oncogenes. Cell. 1984 May;37(1):9–20. doi: 10.1016/0092-8674(84)90296-4. [DOI] [PubMed] [Google Scholar]
- Heslop J. P., Blakeley D. M., Brown K. D., Irvine R. F., Berridge M. J. Effects of bombesin and insulin on inositol (1,4,5)trisphosphate and inositol (1,3,4)trisphosphate formation in Swiss 3T3 cells. Cell. 1986 Dec 5;47(5):703–709. doi: 10.1016/0092-8674(86)90513-1. [DOI] [PubMed] [Google Scholar]
- Isacke C. M., Meisenhelder J., Brown K. D., Gould K. L., Gould S. J., Hunter T. Early phosphorylation events following the treatment of Swiss 3T3 cells with bombesin and the mammalian bombesin-related peptide, gastrin-releasing peptide. EMBO J. 1986 Nov;5(11):2889–2898. doi: 10.1002/j.1460-2075.1986.tb04584.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- James R., Bradshaw R. A. Polypeptide growth factors. Annu Rev Biochem. 1984;53:259–292. doi: 10.1146/annurev.bi.53.070184.001355. [DOI] [PubMed] [Google Scholar]
- Jensen R. T., Jones S. W., Folkers K., Gardner J. D. A synthetic peptide that is a bombesin receptor antagonist. Nature. 1984 May 3;309(5963):61–63. doi: 10.1038/309061a0. [DOI] [PubMed] [Google Scholar]
- King A. C., Cuatrecasas P. Peptide hormone-induced receptor mobility, aggregation, and internalization. N Engl J Med. 1981 Jul 9;305(2):77–88. doi: 10.1056/NEJM198107093050206. [DOI] [PubMed] [Google Scholar]
- Letterio J. J., Coughlin S. R., Williams L. T. Pertussis toxin-sensitive pathway in the stimulation of c-myc expression and DNA synthesis by bombesin. Science. 1986 Nov 28;234(4780):1117–1119. doi: 10.1126/science.3465038. [DOI] [PubMed] [Google Scholar]
- Lin C. R., Chen W. S., Lazar C. S., Carpenter C. D., Gill G. N., Evans R. M., Rosenfeld M. G. Protein kinase C phosphorylation at Thr 654 of the unoccupied EGF receptor and EGF binding regulate functional receptor loss by independent mechanisms. Cell. 1986 Mar 28;44(6):839–848. doi: 10.1016/0092-8674(86)90006-1. [DOI] [PubMed] [Google Scholar]
- Lopez-Rivas A., Mendoza S. A., Nånberg E., Sinnett-Smith J., Rozengurt E. Ca2+-mobilizing actions of platelet-derived growth factor differ from those of bombesin and vasopressin in Swiss 3T3 mouse cells. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5768–5772. doi: 10.1073/pnas.84.16.5768. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Masu Y., Nakayama K., Tamaki H., Harada Y., Kuno M., Nakanishi S. cDNA cloning of bovine substance-K receptor through oocyte expression system. 1987 Oct 29-Nov 4Nature. 329(6142):836–838. doi: 10.1038/329836a0. [DOI] [PubMed] [Google Scholar]
- McDonald T. J., Jörnvall H., Nilsson G., Vagne M., Ghatei M., Bloom S. R., Mutt V. Characterization of a gastrin releasing peptide from porcine non-antral gastric tissue. Biochem Biophys Res Commun. 1979 Sep 12;90(1):227–233. doi: 10.1016/0006-291x(79)91614-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Minamino N., Kangawa K., Matsuo H. Neuromedin B: a novel bombesin-like peptide identified in porcine spinal cord. Biochem Biophys Res Commun. 1983 Jul 29;114(2):541–548. doi: 10.1016/0006-291x(83)90814-8. [DOI] [PubMed] [Google Scholar]
- Minamino N., Kangawa K., Matsuo H. Neuromedin C: a bombesin-like peptide identified in porcine spinal cord. Biochem Biophys Res Commun. 1984 Feb 29;119(1):14–20. doi: 10.1016/0006-291x(84)91611-5. [DOI] [PubMed] [Google Scholar]
- Minamino N., Sudoh T., Kangawa K., Matsuo H. Neuromedin B-32 and B-30: two "big" neuromedin B identified in porcine brain and spinal cord. Biochem Biophys Res Commun. 1985 Jul 31;130(2):685–691. doi: 10.1016/0006-291x(85)90471-1. [DOI] [PubMed] [Google Scholar]
- Moody T. W., Pert C. B., Gazdar A. F., Carney D. N., Minna J. D. High levels of intracellular bombesin characterize human small-cell lung carcinoma. Science. 1981 Dec 11;214(4526):1246–1248. doi: 10.1126/science.6272398. [DOI] [PubMed] [Google Scholar]
- Muir J. G., Murray A. W. Bombesin and phorbol ester stimulate phosphatidylcholine hydrolysis by phospholipase C: evidence for a role of protein kinase C. J Cell Physiol. 1987 Mar;130(3):382–391. doi: 10.1002/jcp.1041300311. [DOI] [PubMed] [Google Scholar]
- 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]
- Palumbo A. P., Rossino P., Comoglio P. M. Bombesin stimulation of c-fos and c-myc gene expression in cultures of Swiss 3T3 cells. Exp Cell Res. 1986 Nov;167(1):276–280. doi: 10.1016/0014-4827(86)90226-0. [DOI] [PubMed] [Google Scholar]
- Price J., Penman E., Bourne G. L., Rees L. H. Characterisation of bombesin-like immunoreactivity in human fetal lung. Regul Pept. 1983 Dec;7(4):315–322. doi: 10.1016/0167-0115(83)90103-9. [DOI] [PubMed] [Google Scholar]
- Rodriguez-Pena A., Rozengurt E. Disappearance of Ca2+-sensitive, phospholipid-dependent protein kinase activity in phorbol ester-treated 3T3 cells. Biochem Biophys Res Commun. 1984 May 16;120(3):1053–1059. doi: 10.1016/s0006-291x(84)80213-2. [DOI] [PubMed] [Google Scholar]
- Rodriguez-Pena A., Rozengurt E. Phosphorylation of an acidic mol. wt. 80 000 cellular protein in a cell-free system and intact Swiss 3T3 cells: a specific marker of protein kinase C activity. EMBO J. 1986 Jan;5(1):77–83. doi: 10.1002/j.1460-2075.1986.tb04180.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rodriguez-Pena A., Rozengurt E. Serum, like phorbol esters, rapidly activates protein kinase C in intact quiescent fibroblasts. EMBO J. 1985 Jan;4(1):71–76. doi: 10.1002/j.1460-2075.1985.tb02319.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenblatt M. Peptide hormone antagonists that are effective in vivo. Lessons from parathyroid hormone. N Engl J Med. 1986 Oct 16;315(16):1004–1013. doi: 10.1056/NEJM198610163151606. [DOI] [PubMed] [Google Scholar]
- Rozengurt E. Early signals in the mitogenic response. Science. 1986 Oct 10;234(4773):161–166. doi: 10.1126/science.3018928. [DOI] [PubMed] [Google Scholar]
- Rozengurt E. Growth factors, cell proliferation and cancer: an overview. Mol Biol Med. 1983 Jul;1(1):169–181. [PubMed] [Google Scholar]
- Rozengurt E., Murray M., Zachary I., Collins M. Protein kinase C activation enhances cAMP accumulation in Swiss 3T3 cells: inhibition by pertussis toxin. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2282–2286. doi: 10.1073/pnas.84.8.2282. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rozengurt E., Rodriguez-Pena A., Coombs M., Sinnett-Smith J. Diacylglycerol stimulates DNA synthesis and cell division in mouse 3T3 cells: role of Ca2+-sensitive phospholipid-dependent protein kinase. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5748–5752. doi: 10.1073/pnas.81.18.5748. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rozengurt E., Rodriguez-Pena M., Smith K. A. Phorbol esters, phospholipase C, and growth factors rapidly stimulate the phosphorylation of a Mr 80,000 protein in intact quiescent 3T3 cells. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7244–7248. doi: 10.1073/pnas.80.23.7244. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rozengurt E., Sinnett-Smith J. W. Bombesin induction of c-fos and c-myc proto-oncogenes in Swiss 3T3 cells: significance for the mitogenic response. J Cell Physiol. 1987 May;131(2):218–225. doi: 10.1002/jcp.1041310211. [DOI] [PubMed] [Google Scholar]
- 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]
- Sausville E. A., Lebacq-Verheyden A. M., Spindel E. R., Cuttitta F., Gazdar A. F., Battey J. F. Expression of the gastrin-releasing peptide gene in human small cell lung cancer. Evidence for alternative processing resulting in three distinct mRNAs. J Biol Chem. 1986 Feb 15;261(5):2451–2457. [PubMed] [Google Scholar]
- Spindel E. R., Chin W. W., Price J., Rees L. H., Besser G. M., Habener J. F. Cloning and characterization of cDNAs encoding human gastrin-releasing peptide. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5699–5703. doi: 10.1073/pnas.81.18.5699. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spindel E. R., Sunday M. E., Hofler H., Wolfe H. J., Habener J. F., Chin W. W. Transient elevation of messenger RNA encoding gastrin-releasing peptide, a putative pulmonary growth factor in human fetal lung. J Clin Invest. 1987 Oct;80(4):1172–1179. doi: 10.1172/JCI113176. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sporn M. B., Roberts A. B. Peptide growth factors and inflammation, tissue repair, and cancer. J Clin Invest. 1986 Aug;78(2):329–332. doi: 10.1172/JCI112580. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stabel S., Rodriguez-Pena A., Young S., Rozengurt E., Parker P. J. Quantitation of protein kinase C by immunoblot--expression in different cell lines and response to phorbol esters. J Cell Physiol. 1987 Jan;130(1):111–117. doi: 10.1002/jcp.1041300116. [DOI] [PubMed] [Google Scholar]
- Suzuki M., Yamaguchi K., Abe K., Adachi N., Nagasaki K., Asanuma F., Adachi I., Kimura S., Terada M., Taya Y. Detection of gastrin-releasing peptide mRNA in small cell lung carcinomas and medullary thyroid carcinomas using synthetic oligodeoxyribonucleotide probes. Jpn J Clin Oncol. 1987 Jun;17(2):157–163. [PubMed] [Google Scholar]
- 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]
- Vara F., Schneider J. A., Rozengurt E. Ionic responses rapidly elicited by activation of protein kinase C in quiescent Swiss 3T3 cells. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2384–2388. doi: 10.1073/pnas.82.8.2384. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wakshull E. M., Wharton W. Stabilized complexes of epidermal growth factor and its receptor on the cell surface stimulate RNA synthesis but not mitogenesis. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8513–8517. doi: 10.1073/pnas.82.24.8513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weber S., Zuckerman J. E., Bostwick D. G., Bensch K. G., Sikic B. I., Raffin T. A. Gastrin releasing peptide is a selective mitogen for small cell lung carcinoma in vitro. J Clin Invest. 1985 Jan;75(1):306–309. doi: 10.1172/JCI111690. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinstein I. B. Growth factors, oncogenes, and multistage carcinogenesis. J Cell Biochem. 1987 Mar;33(3):213–224. doi: 10.1002/jcb.240330308. [DOI] [PubMed] [Google Scholar]
- Wharton J., Polak J. M., Bloom S. R., Ghatei M. A., Solcia E., Brown M. R., Pearse A. G. Bombesin-like immunoreactivity in the lung. Nature. 1978 Jun 29;273(5665):769–770. doi: 10.1038/273769a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Wood S. M., Wood J. R., Ghatei M. A., Lee Y. C., O'Shaughnessy D., Bloom S. R. Bombesin, somatostatin and neurotensin-like immunoreactivity in bronchial carcinoma. J Clin Endocrinol Metab. 1981 Dec;53(6):1310–1312. doi: 10.1210/jcem-53-6-1310. [DOI] [PubMed] [Google Scholar]
- Yamaguchi K., Abe K., Kameya T., Adachi I., Taguchi S., Otsubo K., Yanaihara N. Production and molecular size heterogeneity of immunoreactive gastrin-releasing peptide in fetal and adult lungs and primary lung tumors. Cancer Res. 1983 Aug;43(8):3932–3939. [PubMed] [Google Scholar]
- Zachary I., Millar J., Nånberg E., Higgins T., Rozengurt E. Inhibition of bombesin-induced mitogenesis by pertussis toxin: dissociation from phospholipase C pathway. Biochem Biophys Res Commun. 1987 Jul 31;146(2):456–463. doi: 10.1016/0006-291x(87)90551-1. [DOI] [PubMed] [Google Scholar]
- Zachary I., Rozengurt E. A substance P antagonist also inhibits specific binding and mitogenic effects of vasopressin and bombesin-related peptides in Swiss 3T3 cells. Biochem Biophys Res Commun. 1986 May 29;137(1):135–141. doi: 10.1016/0006-291x(86)91186-1. [DOI] [PubMed] [Google Scholar]
- Zachary I., Rozengurt E. High-affinity receptors for peptides of the bombesin family in Swiss 3T3 cells. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7616–7620. doi: 10.1073/pnas.82.22.7616. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zachary I., Rozengurt E. Identification of a receptor for peptides of the bombesin family in Swiss 3T3 cells by affinity cross-linking. J Biol Chem. 1987 Mar 25;262(9):3947–3950. [PubMed] [Google Scholar]
- Zachary I., Rozengurt E. Internalization and degradation of peptides of the bombesin family in Swiss 3T3 cells occurs without ligand-induced receptor down-regulation. EMBO J. 1987 Aug;6(8):2233–2239. doi: 10.1002/j.1460-2075.1987.tb02495.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zachary I., Rozengurt E. Modulation of the epidermal growth factor receptor by mitogenic ligands: effects of bombesin and role of protein kinase C. Cancer Surv. 1985;4(4):729–765. [PubMed] [Google Scholar]
- Zachary I., Woll P. J., Rozengurt E. A role for neuropeptides in the control of cell proliferation. Dev Biol. 1987 Dec;124(2):295–308. doi: 10.1016/0012-1606(87)90483-0. [DOI] [PubMed] [Google Scholar]
