SpringerProtocols: Abstract: Cancer Cell Receptor Internalization and Proliferation: Effects of Neuropeptide Analogs

Cancer Cell Receptor Internalization and Proliferation: Effects of Neuropeptide Analogs

By: Terry W. Moody², Michael Schumann³, Robert T. Jensen⁴

Abstract

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Neuropeptide receptors can be used as molecular targets to deliver chemotherapeutic drugs into cancer cells. The gastrin-releasing peptide (GRP) receptor was used to deliver a camptothecin (CPT)–bombesin (BB) conjugate into the lung cancer cell line NCI-H1299. The CPT–BB conjugate was metabolized by NCI-H1299 intracellular enzymes releasing the cytotoxic CPT. Receptor binding methods and fluorescent methods are presented for studying the internalization of GRP receptors. The MTT, clonogenic, and ³H-thymidine uptake assays are presented for studying the proliferation of NCI-H1299 cells. The results indicate that CPT-BB conjugates are internalized as a result of receptor-mediated endocytosis.

Affiliation(s): (2) Department of Health and Human Services, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, USA
(3) Digestive Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD, USA
(4) Digestive Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD, USA

Book Title: Neuropeptide Techniques

Series: Neuromethods | Volume: 39 | Pub. Date: Nov-13-2007 | Page Range: 115-129 | DOI: 10.1007/978-1-60327-099-1_9

Subject: Neuroscience

References

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1.	Moody, T.W., O’Donohue, T.L., and Jacobowitz, D.M. Biochemical localization and characterization of bombesin-like peptides in discrete regions of rat brain. Peptides, 1981; 2:75–79.

2.	Moody, T.W., Pert, C.B., Rivier, J., and Brown, M.R. Bombesin: Specific binding to rat brain membranes. Proc. Natl. Acad. Sci. USA, 1978; 75: 5372–5376.

3.	Moody, T.W., Thoa, N.B., O’Donohue, T.L., and Pert, C.B. Bombesin-like peptides in rat brain: Localization in synaptosomes and release from hypothalamic slices. Life Sci., 1980; 26: 1707–1712.

4.	Gibbs, J., Fauser, D.J., Rowe, E.A., Rolls, B.J., Rolls, E.T., and Maddison, S.P. Bombesin suppresses feeding in rats. Nature, 1979; 282: 208–210.

5.	Anastasi, A., Erspamer, V., and Bucci, M. Isolation and amino acid sequences of alytesin and bombesin: Two analogous active tetradecapeptides from the skin of European discoglossid frogs. Arch. Biochem. Biophys., 1973; 148: 443–446.

6.	Brown, M.R., Rivier, J., and Vale, W. Bombesin: Potent effects on thermoregulation in the rat. Science, 1977; 196: 998–1000.

7.	Brown, M., Rivier, J., and Vale, W. Bombesin affects the central nervous system to produce hyperglycemia in rats. Life Sci., 1978; 21: 1729–1734.

8.	Merali, Z., Moody, T., Kateb, P., and Piggins, H. Antagonism of satiety and grooming effects of bombesin by antiserum to bombesin and by [Tyr⁴, D-Phe¹²]bombesin: Central versus peripheral effects. Ann. NY Acad. Sci., 1988; 547: 489–492.

9.	Tache, Y. CNS peptides and regulation of gastric acid secretion. Ann. Rev. Physiol., 1988; 50: 19–39.

10.	Merali, Z., Kent, P., and Anisman, H. Role of bombesin-related peptides in the mediation or integration of the stress response. Life Sci., 2002; 59: 272–287.

11.	Cuttitta, F., Carney, D.N., Mulshine, J., Moody, T.W., Fedorko, J., Fischler, A., and Minna, J.D. Bombesin-like peptides can function as autocrine growth factors in human small cell lung cancer. Nature, 1985; 316:823–825.

12.	Korman, L.Y., Carney, D.N., Citron, M.L., and Moody, T.W. Secretin/VIP stimulated secretion of bombesin-like peptides from human small cell lung cancer. Cancer Res., 1986; 46: 1214–1218.

13.	Moody, T.W., Mahmoud, S., Staley, J., Naldini, L., Cirillo, D., South, V., Felder, S., and Kris, R. Human glioblastoma cell lines have neuropeptide receptors for bombesin/GRP. J. Mol. Neurosci., 1989; 1: 235–242.

14.	Benya, R.V., Kusui, T., Pradhan, T.K., Battey, J.F., and Jensen, R.T. Expression and characterization of cloned human bombesin receptors. Mol. Pharmacol., 1995; 47: 10–20.

15.	Carney, D.N., Cuttitta, F., Moody, T.W., and Minna, J.D. Selective stimulation of small cell lung cancer clonal growth by bombesin and gastrin releasing peptide. Cancer Res., 1987; 47: 821–825.

16.	Koh, S.W., Leyton, J., and Moody, T.W. Bombesin activates MAP kinase in non-small cell lung cancer cells. Peptides, 1999; 20: 121–126.

17.	Mahmoud, S., Staley, J., Taylor, J., Bogden, A., Moreau, J.P., Coy, D., Avis, I., Cuttitta, F., Mulshine, J.L., and Moody, T.W. [Psi ^{13, 14}] bombesin analogues inhibit growth of small cell lung cancer in vitro and in vivo. Cancer Res., 1991; 51: 1798–1802.

18.	Moody, T.W., Zia, F., Venugopal, R., Patierno, S., LeBan, J., and McDermod, J. BW2258: A GRP receptor antagonist which inhibits small cell lung cancer growth. Life Sci., 1995; 56: 523–529.

19.	Moody, T.W., Leyton, J., Garcia, L., and Jensen, R.T. Nonpeptide gastrin releasing peptide receptor antagonists inhibit the proliferation of lung cancer cells. Eur. J. Pharmacol., 2003b; 474: 21–29.

20.	McDonald, T.J., Jornvall, J., Nilsson, G., Vagne, M., Ghatei, M., Bloom, S.R., and Mutt, V. Characterization of a gastrin-releasing peptide from porcine non-antral gastric tissue. Biochem. Biophys. Res. Comm., 1979; 90: 227–233.

21.	Minamino, N., Kangawa, K., and Matsuo, H. Neuromedin B: A novel bombesin-like peptide identified in porcine spinal cord. Biochem. Biophys. Res. Comm., 1983; 114: 541–548.

22.	Benya, R.V., Wada, E., Battey, J.F., Fathi, Z., Wang, L.H., Mantey, S.A., Coy, D.H., and Jensen, R.T. Neuromedin B receptors retain functional expression when transfected into BALB 3T3 fibroblasts: Analysis of binding, kinetics, stoichiometry, modulation by guanine nucleotide-binding proteins, and signal transduction and comparison with natively expressed receptors. Mol. Pharmacol., 1992; 42: 1058–1068.

23.	Benya, R.V., Fathi, Z., Pradhan, T., Battey, J.F., Kusui, T., and Jensen, R.T. Gastrin-releasing peptide receptor-induced internalization, down-regulation, desensitization and growth: Possible role of cAMP. Mol. Pharmacol., 1994; 46: 235–245.

24.	Battey, J.F., Way, J.M., Corjay, M.H., Shapira, H., Kusano, K., Harkins, R., Wu, J.M., Slattery, T., Mann, E., and Feldman, R.I. Molecular cloning of the bombesin/gastrin-releasing peptide receptor from Swiss 3T3 cells. Proc. Natl. Acad. Sci. USA, 1991; 88: 395–399.

25.	Wada, E., Way, J., Shapira, H., Kusamo, K., Lebacq-Verheyden, A.M., Coy, D., Jensen, R.T., and Battey, J. cDNA cloning, characterization and brain region-specific expression of a neuromedin-B preferring bombesin receptor. Neuron, 1991; 6: 421–430.

26.	Fathi, Z., Corjay, M.H., Shapira, H., Wada, E., Benya, R., Jensen, R., Viallet, J., Sausville, E.A., and Battey, J.F. BRS-3: Novel bombesin receptor subtype selectively expressed in testis and lung carcinoma cells. J. Biol. Chem., 1993; 268: 5979–5984.

27.	Ryan, R.R., Weber, H.C., Mantey, S.A., Hou, W., Hilburger, M.E., Pradhan, T.K., Coy, D.H., and Jensen, R.T. Pharmacology and intracellular signaling mechanisms of the native human orphan receptor BRS-3 in lung cancer cells. J. Pharmacol. Exp. Ther., 1998a; 287: 366–380.

28.	Ryan, R.R., Weber, H.C., Hou, W., Sainz, E., Mantey, S.A., Battey, J.F., Coy, D.H., and Jensen, R.T. Ability of various bombesin receptor agonists and antagonists to alter intracellular signaling of the human orphan receptor BRS-3. J. Biol. Chem., 1998b; 273: 13613–13624.

29.	Mantey, S.A., Weber, H.C., Sainz, E., Akeson, M., Ryan, R.R., Pradhan, T.K., Searles, R.P., Spindel, E.R., Battey, J.F., Coy, D.H., and Jensen, R.T. Discovery of a high affinity radioligand for the human orphan receptor, bombesin receptor subtype 3, which demonstrates that it has a unique pharmacology compared with other mammalian bombesin receptors. J. Biol. Chem., 1997; 272: 26062–26071.

30.	Pradhan, T.K., Katsuno, T., Taylor, J.E., Kim, S.H., Ryan, R.R., Mantey, S.A., Donohue, P.J., Weber, H.C., Sainz, E., Battey, J.F., Coy, D.H., and Jensen, R.T. Identification of a unique ligand which has high affinity for all four bombesin receptor subtypes. Eur. J. Pharmacol., 1998; 343: 275–287.

31.	Reubi, J.C., Wenger, S., Schumuckli-Maurer, J., Schaer, J.C., and Gugger, M. Bombesin receptor subtypes in human cancers: Detection with the universal radoligand (125)I-[D-TYR(6), beta-ALA(11),PHE(13), NLE(14)] bombesin(6–14). Clin. Cancer Res., 2002; 8: 1139–1146.

32.	Reubi, J.C. Peptide receptors as molecular targets for cancer diagnosis and therapy. Endocr. Rev., 2003; 24: 389–427.

33.	Moody, T.W., Chan, D., Fahrenkrug, J., and Jensen, R.T. Neuropeptides as autocrine growth factors in cancer cells. Curr. Pharm. Des., 2003a; 9: 495–509.

34.	Schally, A.V., and Nagy, A. New approaches to treatment of various cancers based on cytotoxic analogs of LHRH, somatostatin and bombesin. Life Sci., 2003; 72: 2305–2320.

35.	Szereday, Z., Schally, A.V., Nagy, A., Plonowski, A., Bajo, A.M., Halmos, G., Szepeshazi, K., and Groot, K. Effective treatment of experimental U-87MG human glioblastoma in nude mice with a targeted cytotoxic bombesin analogue, AN-215. Br. J. Cancer, 2002; 86: 1322–1327.

36.	Moody, T.W., Mantey, S.A., Pradhan, T., Schumann, R., Nakagawa, A., Martinez, A., Fusilier, J., Coy, D.H., and Jensen, R.T. Development of high affinity camptothecin-bombesin conjugates which have targeted cytotoxicity for bombesin receptor-containing cells. J. Biol. Chem., 2004; in publication.

37.	Moody, T.W., Zia, F., Venugopal, R., Fagarasan, M., Oie, H., and Hu, V. GRP receptors are present in non small cell lung cancer cells. J. Cell. Biochem. Suppl., 1996; 24: 247–256.

38.	Fuselier, J.A., Sun, L., Woltering, S.N., Murphy, W.A., Vasilevich, N., and Coy, D.H. An adjustable release rate linking strategy for cytotoxic peptide conjugates. Bioorg. Med. Chem. Lett., 2003; 13: 799–803.

39.	Cheng, Y., and Prusoff, W.H. Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50 percent inhibition (I₅₀) of an enzymatic reaction. Biochem. Pharmacol., 1973; 22: 3099–3108.

40.	Marchese, A., Chen, C., Kim, Y.M., and Benovic, J.L. The ins and outs of G protein-coupled receptor trafficking. Trends Biochem. Sci., 2003; 28: 369–376.

41.	Grady, E.F., Slice, L.W., Brant, W.O., Walsh, J.H., Payan, D.G., and Bunnett, N.W. Direct observation of endocytosis of gastrin releasing peptide and its receptor. J. Biol. Chem., 1995; 270: 4603–4611.

42.	Slice, L.W., Yee, H.F., Jr., and Walsh, J.H. Visualization of internalization and recycling of the gastrin releasing peptide receptor-green fluorescent protein chimera expressed in epithelial cells. Receptors Channels, 1998; 6: 201–212.

43.	Heuser, J.E., and Anderson, R.G. Hypertonic media inhibit receptor-mediated endocytosis by blocking clathrin-coated pit formation. J. Cell Biol., 1989; 108: 389–400.

44.	Wang, L.H., Rothberg, K.G., and Anderson, R.G. Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J. Cell. Biol., 1993; 123: 1107–1117.

45.	Schiller, J.H., Kim, K., Hutson, P., DeVore, R., Glick, J., Stewart, J., and Johnson, D. Phase II study of topotecan in patients with extensive stage small-cell carcinoma of the lung: An Eastern Cooperative Oncology Group Trial. J. Clin. Oncol., 1996; 14: 2345–2352.

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