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Suppression of the proliferation of human U-87 MG glioblastoma cells by new antagonists of growth hormone-releasing hormone in vivo and in vitro

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Abstract

Five-year survival of patients afflicted with glioblastoma multiforme (GBM) is rare, making this cancer one of the most feared malignancies. Previously, we reported that growth hormone-releasing hormone (GHRH) is a potent growth factor in cancers. The present work evaluated the effects of two antagonistic analogs of GHRH (MIA-604 and MIA-690) on the proliferation of U-87 MG GBM tumors, in vivo as well as in vitro. Both analogs were administered subcutaneously and dose-dependently inhibited the growth of tumors transplanted into nude mice (127 animals in seven groups). The analogs also inhibited cell proliferation in vitro, decreased cell size, and promoted apoptotic and autophagic processes. Both antagonists stimulated contact inhibition, as indicated by the expression of the E-cadherin–β-catenin complex and integrins, and decreased the release of humoral regulators of glial growth such as FGF, PDGFβ, and TGFβ, as revealed by genomic or proteomic detection methods. The GHRH analogs downregulated other tumor markers (Jun-proto-oncogene, mitogen-activated protein kinase-1, and melanoma cell adhesion molecule), upregulated tumor suppressors (p53, metastasis suppressor-1, nexin, TNF receptor 1A, BCL-2-associated agonist of cell death, and ifκBα), and inhibited the expression of the regulators of angiogenesis and invasion (angiopoetin-1, VEGF, matrix metallopeptidase-1, S100 calcium binding protein A4, and synuclein-γ). Our findings indicate that GHRH antagonists inhibit growth of GBMs by multiple mechanisms and decrease both tumor cell size and number.

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References

  1. Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352(10):987–996. doi:10.1056/NEJMoa043330

    Article  PubMed  CAS  Google Scholar 

  2. Barabutis N, Schally AV (2010) Growth hormone-releasing hormone: extrapituitary effects in physiology and pathology. Cell Cycle 9(20):4110–4116

    Article  PubMed  CAS  Google Scholar 

  3. Engel JB, Schally AV (2007) Drug Insight: clinical use of agonists and antagonists of luteinizing-hormone-releasing hormone. Nat Clin Pract Endocrinol Metab 3(2):157–167. doi:10.1038/ncpendmet0399

    Article  PubMed  CAS  Google Scholar 

  4. Schally AV, Varga JL, Engel JB (2008) Antagonists of growth-hormone-releasing hormone: an emerging new therapy for cancer. Nat Clin Pract Endocrinol Metab 4(1):33–43. doi:10.1038/ncpendmet0677

    Article  PubMed  CAS  Google Scholar 

  5. Schally AV, Nagy A (2003) New approaches to treatment of various cancers based on cytotoxic analogs of LHRH, somatostatin and bombesin. Life Sci 72(21):2305–2320

    Article  PubMed  CAS  Google Scholar 

  6. Schally AV, Comaru-Schally AM, Nagy A, Kovacs M, Szepeshazi K, Plonowski A, Varga JL, Halmos G (2001) Hypothalamic hormones and cancer. Front Neuroendocrinol 22(4):248–291. doi:10.1006/frne.2001.0217

    Article  PubMed  CAS  Google Scholar 

  7. Rick FG, Buchholz S, Schally AV, Szalontay L, Krishan A, Datz C, Stadlmayr A, Aigner E, Perez R, Seitz S, Block NL, Hohla F (2012) Combination of gastrin-releasing peptide antagonist with cytotoxic agents produces synergistic inhibition of growth of human experimental colon cancers. Cell Cycle 11(13):2518–2525. doi:10.4161/cc.20900

    Article  PubMed  CAS  Google Scholar 

  8. Hohla F, Buchholz S, Schally AV, Krishan A, Rick FG, Szalontay L, Papadia A, Halmos G, Koster F, Aigner E, Datz C, Seitz S (2010) Targeted cytotoxic somatostatin analog AN-162 inhibits growth of human colon carcinomas and increases sensitivity of doxorubicin resistant murine leukemia cells. Cancer Lett 294(1):35–42. doi:10.1016/j.canlet.2010.01.018

    Article  PubMed  CAS  Google Scholar 

  9. Treszl A, Schally AV, Seitz S, Szalontay L, Rick FG, Szepeshazi K, Halmos G (2009) Inhibition of human non-small cell lung cancers with a targeted cytotoxic somatostatin analog, AN-162. Peptides 30(9):1643–1650. doi:10.1016/j.peptides.2009.06.007

    Article  PubMed  CAS  Google Scholar 

  10. Szepeshazi K, Schally AV, Rick FG, Block NL, Vidaurre I, Halmos G, Szalontay L (2012) Powerful inhibition of in-vivo growth of experimental hepatic cancers by bombesin/gastrin-releasing peptide antagonist RC-3940-II. Anticancer Drugs 23(9):906–913. doi:10.1097/CAD.0b013e328354bd25

    PubMed  CAS  Google Scholar 

  11. Rick FG, Schally AV, Block NL, Nadji M, Szepeshazi K, Zarandi M, Vidaurre I, Perez R, Halmos G, Szalontay L (2011) Antagonists of growth hormone-releasing hormone (GHRH) reduce prostate size in experimental benign prostatic hyperplasia. Proc Natl Acad Sci U S A 108(9):3755–3760. doi:10.1073/pnas.1018086108

    Article  PubMed  CAS  Google Scholar 

  12. Rick FG, Szalontay L, Schally AV, Block NL, Nadji M, Szepeshazi K, Vidaurre I, Zarandi M, Kovacs M, Rekasi Z (2012) Combining growth hormone-releasing hormone antagonist with luteinizing hormone-releasing hormone antagonist greatly augments benign prostatic hyperplasia shrinkage. J Urol 187(4):1498–1504. doi:10.1016/j.juro.2011.11.081

    Article  PubMed  CAS  Google Scholar 

  13. Pozsgai E, Schally AV, Zarandi M, Varga JL, Vidaurre I, Bellyei S (2010) The effect of GHRH antagonists on human glioblastomas and their mechanism of action. Int J Cancer 127(10):2313–2322. doi:10.1002/ijc.25259

    Article  PubMed  CAS  Google Scholar 

  14. Garcia-Fernandez MO, Schally AV, Varga JL, Groot K, Busto R (2003) The expression of growth hormone-releasing hormone (GHRH) and its receptor splice variants in human breast cancer lines; the evaluation of signaling mechanisms in the stimulation of cell proliferation. Breast Cancer Res Treat 77(1):15–26

    Article  PubMed  CAS  Google Scholar 

  15. Klukovits A, Schally AV, Szalontay L, Vidaurre I, Papadia A, Zarandi M, Varga JL, Block NL, Halmos G (2012) Novel antagonists of growth hormone-releasing hormone inhibit growth and vascularization of human experimental ovarian cancers. Cancer 118(3):670–680. doi:10.1002/cncr.26291

    Article  PubMed  CAS  Google Scholar 

  16. Rick FG, Schally AV, Szalontay L, Block NL, Szepeshazi K, Nadji M, Zarandi M, Hohla F, Buchholz S, Seitz S (2012) Antagonists of growth hormone-releasing hormone inhibit growth of androgen-independent prostate cancer through inactivation of ERK and Akt kinases. Proc Natl Acad Sci U S A 109(5):1655–1660. doi:10.1073/pnas.1120588109

    Article  PubMed  CAS  Google Scholar 

  17. Pozsgai E, Schally AV, Hocsak E, Zarandi M, Rick F, Bellyei S (2011) The effect of a novel antagonist of growth hormone releasing hormone on cell proliferation and on the key cell signaling pathways in nine different breast cancer cell lines. Int J Oncol 39(4):1025–1032. doi:10.3892/ijo.2011.1098

    PubMed  CAS  Google Scholar 

  18. Perez R, Schally AV, Vidaurre I, Rincon R, Block NL, Rick FG (2012) Antagonists of growth hormone-releasing hormone suppress in vivo tumor growth and gene expression in triple negative breast cancers. Oncotarget 3(9):988–997

    PubMed  Google Scholar 

  19. Papadia A, Schally AV, Halmos G, Varga JL, Seitz S, Buchholz S, Rick F, Zarandi M, Bellyei S, Treszl A, Szalontay L, Lucci JA (2011) Growth hormone-releasing hormone antagonists inhibit growth of human ovarian cancer. Horm Metab Res 43(11):816–820. doi:10.1055/s-0031-1287766

    Article  PubMed  CAS  Google Scholar 

  20. Jaszberenyi M, Rick FG, Szalontay L, Block NL, Zarandi M, Cai RZ, Schally AV (2012) Beneficial effects of novel antagonists of GHRH in different models of Alzheimer’s disease. Aging 4(11):755–767

    PubMed  CAS  Google Scholar 

  21. Mareel M, Leroy A (2003) Clinical, cellular, and molecular aspects of cancer invasion. Physiol Rev 83(2):337–376. doi:10.1152/physrev.00024.2002

    PubMed  CAS  Google Scholar 

  22. Bredesen DE, Rao RV, Mehlen P (2006) Cell death in the nervous system. Nature 443(7113):796–802. doi:10.1038/nature05293

    Article  PubMed  CAS  Google Scholar 

  23. Notte A, Leclere L, Michiels C (2011) Autophagy as a mediator of chemotherapy-induced cell death in cancer. Biochem Pharmacol 82(5):427–434. doi:10.1016/j.bcp. 2011.06.015

    Article  PubMed  CAS  Google Scholar 

  24. Goel S, Duda DG, Xu L, Munn LL, Boucher Y, Fukumura D, Jain RK (2011) Normalization of the vasculature for treatment of cancer and other diseases. Physiol Rev 91(3):1071–1121. doi:10.1152/physrev.00038.2010

    Article  PubMed  CAS  Google Scholar 

  25. Kiaris H, Schally AV, Varga JL, Groot K, Armatis P (1999) Growth hormone-releasing hormone: an autocrine growth factor for small cell lung carcinoma. Proc Natl Acad Sci U S A 96(26):14894–14898

    Article  PubMed  CAS  Google Scholar 

  26. Kiaris H, Schally AV, Varga JL (2000) Antagonists of growth hormone-releasing hormone inhibit the growth of U-87MG human glioblastoma in nude mice. Neoplasia 2(3):242–250

    Article  PubMed  CAS  Google Scholar 

  27. Kovacs M, Schally AV, Hohla F, Rick FG, Pozsgai E, Szalontay L, Varga JL, Zarandi M (2010) A correlation of endocrine and anticancer effects of some antagonists of GHRH. Peptides 31(10):1839–1846. doi:10.1016/j.peptides.2010.07.006

    Article  PubMed  CAS  Google Scholar 

  28. Jaeger LB, Banks WA, Varga JL, Schally AV (2005) Antagonists of growth hormone-releasing hormone cross the blood–brain barrier: a potential applicability to treatment of brain tumors. Proc Natl Acad Sci U S A 102(35):12495–12500. doi:10.1073/pnas.0504163102

    Article  PubMed  CAS  Google Scholar 

  29. Ozawa T, James CD (2010) Establishing intracranial brain tumor xenografts with subsequent analysis of tumor growth and response to therapy using bioluminescence imaging. J Vis Exp(41). doi:10.3791/1986

  30. Shankavaram UT, Bredel M, Burgan WE, Carter D, Tofilon P, Camphausen K (2012) Molecular profiling indicates orthotopic xenograft of glioma cell lines simulate a subclass of human glioblastoma. J Cell Mol Med 16(3):545–554. doi:10.1111/j.1582-4934.2011.01345.x

    Article  PubMed  CAS  Google Scholar 

  31. Baumann BC, Dorsey JF, Benci JL, Joh DY, Kao GD (2012) Stereotactic intracranial implantation and in vivo bioluminescent imaging of tumor xenografts in a mouse model system of glioblastoma multiforme. J Vis Exp Sept 25(67). doi:10.3791/4089

  32. Ludwig B, Ziegler CG, Schally AV, Richter C, Steffen A, Jabs N, Funk RH, Brendel MD, Block NL, Ehrhart-Bornstein M, Bornstein SR (2010) Agonist of growth hormone-releasing hormone as a potential effector for survival and proliferation of pancreatic islets. Proc Natl Acad Sci U S A 107(28):12623–12628. doi:10.1073/pnas.1005098107

    Article  PubMed  CAS  Google Scholar 

  33. Kondo Y, Kanzawa T, Sawaya R, Kondo S (2005) The role of autophagy in cancer development and response to therapy. Nat Rev Cancer 5(9):726–734. doi:10.1038/nrc1692

    Article  PubMed  CAS  Google Scholar 

  34. Lefranc F, Kiss R (2006) Autophagy, the Trojan horse to combat glioblastomas. Neurosurg Focus 20(4):E7. doi:10.3171/foc.2006.20.4.4

    Article  PubMed  Google Scholar 

  35. Siejka A, Barabutis N, Schally AV (2011) GHRH antagonist MZ-5-156 increases the expression of AMPK in A549 lung cancer cells. Cell Cycle 10(21):3714–3718. doi:10.4161/cc.10.21.17904

    Article  PubMed  CAS  Google Scholar 

  36. Igney FH, Krammer PH (2002) Death and anti-death: tumour resistance to apoptosis. Nat Rev Cancer 2(4):277–288. doi:10.1038/nrc776

    Article  PubMed  CAS  Google Scholar 

  37. Mizukawa K, Kawamura A, Sasayama T, Tanaka K, Kamei M, Sasaki M, Kohmura E (2006) Synthetic Smac peptide enhances the effect of etoposide-induced apoptosis in human glioblastoma cell lines. J Neurooncol 77(3):247–255. doi:10.1007/s11060-005-9045-5

    Article  PubMed  CAS  Google Scholar 

  38. Jeuken J, van den Broecke C, Gijsen S, Boots-Sprenger S, Wesseling P (2007) RAS/RAF pathway activation in gliomas: the result of copy number gains rather than activating mutations. Acta Neuropathol 114(2):121–133. doi:10.1007/s00401-007-0239-0

    Article  PubMed  CAS  Google Scholar 

  39. Golestaneh N, Mishra B (2005) TGF-beta, neuronal stem cells and glioblastoma. Oncogene 24(37):5722–5730. doi:10.1038/sj.onc.1208925

    Article  PubMed  CAS  Google Scholar 

  40. Galardi S, Mercatelli N, Farace MG, Ciafre SA (2011) NF-kB and c-Jun induce the expression of the oncogenic miR-221 and miR-222 in prostate carcinoma and glioblastoma cells. Nucleic Acids Res 39(9):3892–3902. doi:10.1093/nar/gkr006

    Article  PubMed  CAS  Google Scholar 

  41. Karin M (2006) Nuclear factor-kappaB in cancer development and progression. Nature 441(7092):431–436. doi:10.1038/nature04870

    Article  PubMed  CAS  Google Scholar 

  42. Gyrd-Hansen M, Meier P (2010) IAPs: from caspase inhibitors to modulators of NF-kappaB, inflammation and cancer. Nat Rev Cancer 10(8):561–574. doi:10.1038/nrc2889

    Article  PubMed  CAS  Google Scholar 

  43. Dawson JC, Timpson P, Kalna G, Machesky LM (2011) Mtss1 regulates epidermal growth factor signaling in head and neck squamous carcinoma cells. Oncogene. doi:10.1038/onc.2011.376

  44. Ritchie CK, Giordano A, Khalili K (2000) Integrin involvement in glioblastoma multiforme: possible regulation by NF-kappaB. J Cell Physiol 184(2):214–221. doi:10.1002/1097-4652(200008)184:2<214

    Article  PubMed  CAS  Google Scholar 

  45. Koul D, Shen R, Bergh S, Lu Y, de Groot JF, Liu TJ, Mills GB, Yung WK (2005) Targeting integrin-linked kinase inhibits Akt signaling pathways and decreases tumor progression of human glioblastoma. Mol Cancer Ther 4(11):1681–1688. doi:10.1158/1535-7163.MCT-05-0258

    Article  PubMed  CAS  Google Scholar 

  46. Riemenschneider MJ, Mueller W, Betensky RA, Mohapatra G, Louis DN (2005) In situ analysis of integrin and growth factor receptor signaling pathways in human glioblastomas suggests overlapping relationships with focal adhesion kinase activation. Am J Pathol 167(5):1379–1387. doi:10.1016/S0002-9440(10)61225-4

    Article  PubMed  CAS  Google Scholar 

  47. Engel JB, Keller G, Schally AV, Toller GL, Groot K, Havt A, Armatis P, Zarandi M, Varga JL, Halmos G (2005) Inhibition of growth of experimental human endometrial cancer by an antagonist of growth hormone-releasing hormone. J Clin Endocrinol Metab 90(6):3614–3621. doi:10.1210/jc.2004-2179

    Article  PubMed  CAS  Google Scholar 

  48. Heinrich E, Schally AV, Buchholz S, Rick FG, Halmos G, Mile M, Groot K, Hohla F, Zarandi M, Varga JL (2008) Dose-dependent growth inhibition in vivo of PC-3 prostate cancer with a reduction in tumoral growth factors after therapy with GHRH antagonist MZ-J-7-138. Prostate 68(16):1763–1772. doi:10.1002/pros.20843

    Article  PubMed  CAS  Google Scholar 

  49. Stratmann A, Risau W, Plate KH (1998) Cell type-specific expression of angiopoietin-1 and angiopoietin-2 suggests a role in glioblastoma angiogenesis. Am J Pathol 153(5):1459–1466. doi:10.1016/S0002-9440(10)65733-1

    Article  PubMed  CAS  Google Scholar 

  50. Ouhtit A, Gaur RL, Abd Elmageed ZY, Fernando A, Thouta R, Trappey AK, Abdraboh ME, El-Sayyad HI, Rao P, Raj MG (2009) Towards understanding the mode of action of the multifaceted cell adhesion receptor CD146. Biochim Biophys Acta 1795(2):130–136. doi:10.1016/j.bbcan.2009.01.002

    PubMed  CAS  Google Scholar 

  51. Norden AD, Drappatz J, Wen PY (2009) Antiangiogenic therapies for high-grade glioma. Nat Rev Neurol 5(11):610–620. doi:10.1038/nrneurol.2009.159

    Article  PubMed  CAS  Google Scholar 

  52. Kasza A, Kowanetz M, Poslednik K, Witek B, Kordula T, Koj A (2001) Epidermal growth factor and pro-inflammatory cytokines regulate the expression of components of plasminogen activation system in U373-MG astrocytoma cells. Cytokine 16(5):187–190. doi:10.1006/cyto.2001.0957

    Article  PubMed  CAS  Google Scholar 

  53. Kunapuli P, Kasyapa CS, Hawthorn L, Cowell JK (2004) LGI1, a putative tumor metastasis suppressor gene, controls in vitro invasiveness and expression of matrix metalloproteinases in glioma cells through the ERK1/2 pathway. J Biol Chem 279(22):23151–23157. doi:10.1074/jbc.M314192200

    Article  PubMed  CAS  Google Scholar 

  54. Bellyei S, Schally AV, Zarandi M, Varga JL, Vidaurre I, Pozsgai E (2010) GHRH antagonists reduce the invasive and metastatic potential of human cancer cell lines in vitro. Cancer Lett 293(1):31–40. doi:10.1016/j.canlet.2009.12.014

    Article  PubMed  CAS  Google Scholar 

  55. Zhang H, Kouadio A, Cartledge D, Godwin AK (2011) Role of gamma-synuclein in microtubule regulation. Exp Cell Res 317(10):1330–1339. doi:10.1016/j.yexcr.2010.10.013

    Article  PubMed  CAS  Google Scholar 

  56. Boye K, Maelandsmo GM (2010) S100A4 and metastasis: a small actor playing many roles. Am J Pathol 176(2):528–535. doi:10.2353/ajpath.2010.090526

    Article  PubMed  CAS  Google Scholar 

  57. Wick W, Naumann U, Weller M (2006) Transforming growth factor-beta: a molecular target for the future therapy of glioblastoma. Curr Pharm Des 12(3):341–349

    Article  PubMed  CAS  Google Scholar 

  58. Hohla F, Schally AV, Szepeshazi K, Varga JL, Buchholz S, Koster F, Heinrich E, Halmos G, Rick FG, Kannadka C, Datz C, Kanashiro CA (2006) Synergistic inhibition of growth of lung carcinomas by antagonists of growth hormone-releasing hormone in combination with docetaxel. Proc Natl Acad Sci U S A 103(39):14513–14518. doi:10.1073/pnas.0605309103

    Article  PubMed  CAS  Google Scholar 

  59. Rick FG, Seitz S, Schally AV, Szalontay L, Krishan A, Datz C, Stadlmayr A, Buchholz S, Block NL, Hohla F (2012) GHRH antagonist when combined with cytotoxic agents induces S-phase arrest and additive growth inhibition of human colon cancer. Cell Cycle 11(22):4203–4210

    Article  PubMed  CAS  Google Scholar 

  60. Buchholz S, Schally AV, Engel JB, Hohla F, Heinrich E, Koester F, Varga JL, Halmos G (2007) Potentiation of mammary cancer inhibition by combination of antagonists of growth hormone-releasing hormone with docetaxel. Proc Natl Acad Sci U S A 104(6):1943–1946. doi:10.1073/pnas.0610860104

    Article  PubMed  CAS  Google Scholar 

  61. Varga JL, Schally AV, Horvath JE, Kovacs M, Halmos G, Groot K, Toller GL, Rekasi Z, Zarandi M (2004) Increased activity of antagonists of growth hormone-releasing hormone substituted at positions 8, 9, and 10. Proc Natl Acad Sci U S A 101(6):1708–1713. doi:10.1073/pnas.0307288101

    Article  PubMed  CAS  Google Scholar 

  62. Pozsgai E, Schally AV, Halmos G, Rick F, Bellyei S (2010) The inhibitory effect of a novel cytotoxic somatostatin analogue AN-162 on experimental glioblastoma. Horm Metab Res 42(11):781–786. doi:10.1055/s-0030-1261955

    Article  PubMed  CAS  Google Scholar 

  63. Kletzien RF, Pariza MW, Becker JE, Potter VR (1975) A method using 3-O-methyl-D-glucose and phloretin for the determination of intracellular water space of cells in monolayer culture. Anal Biochem 68(2):537–544

    Article  PubMed  CAS  Google Scholar 

  64. Bender AS, Norenberg MD (1998) Effect of benzodiazepines and neurosteroids on ammonia-induced swelling in cultured astrocytes. J Neurosci Res 54(5):673–680. doi:10.1002/(SICI)1097-4547(19981201)54:5<673

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The work described in this paper was supported by the Medical Research Service of the Veterans Affairs Department, University of Miami, Miller School of Medicine, Departments of Pathology and Medicine, Division of Hematology/Oncology, the South Florida Veterans Affairs Foundation for Research and Education (both to A.V.S.) and the L. Austin Weeks Endowment for Urologic Research (to N.L.B.). This work was supported in part by a grant from the AUA Foundation Research Scholars Program and the AUA Southeastern Section (to F.G. R). Disclosure of potential conflicts of interest: A.V. Schally is listed as co-inventors on the patent applications for GHRH antagonists filed by the University of Miami. However, this article deals with an experimental study on glioblastoma multiforme, which is purely an academic project.

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Jaszberenyi, M., Schally, A.V., Block, N.L. et al. Suppression of the proliferation of human U-87 MG glioblastoma cells by new antagonists of growth hormone-releasing hormone in vivo and in vitro. Targ Oncol 8, 281–290 (2013). https://doi.org/10.1007/s11523-013-0264-y

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