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Published in: Journal of Cancer Research and Clinical Oncology 10/2014

01-10-2014 | Original Article – Cancer Research

Prognosis in human glioblastoma based on expression of ligand growth hormone-releasing hormone, pituitary-type growth hormone-releasing hormone receptor, its splicing variant receptors, EGF receptor and PTEN genes

Authors: Géza Mezey, Andrea Treszl, Andrew V. Schally, Normann L. Block, Laura Vízkeleti, Alíz Juhász, Álmos Klekner, János Nagy, Margit Balázs, Gábor Halmos, László Bognár

Published in: Journal of Cancer Research and Clinical Oncology | Issue 10/2014

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Abstract

Purpose

Glioblastoma (GB) is the most frequent brain tumor. Despite recent improvement in therapeutic strategies, the prognosis of GB remains poor. Growth hormone-releasing hormone (GHRH) may act as a growth factor; antagonists of GHRH have been successfully applied for experimental treatment of different types of tumors. The expression profile of GHRH receptor, its main splice variant SV1 and GHRH have not been investigated in human GB tissue samples.

Methods

We examined the expression of GHRH, full-length pituitary-type GHRH receptor (pGHRHR), its functional splice variant SV1 and non-functional SV2 by RT-PCR in 23 human GB specimens. Epidermal growth factor receptor (EGFR) and phosphatase and tensin homolog gene (PTEN) expression levels were also evaluated by quantitative RT-PCR. Correlations between clinico-pathological parameters and gene expressions were analyzed.

Results

Expression of GHRH was found to be positive in 61.9 % of samples. pGHRH receptor was not expressed in our sample set, while SV1 could be detected in 17.4 % and SV2 in 8.6 % of the GB tissues. In 65.2 and 78.3 % of samples, significant EGFR over-expression or PTEN under-representation could be detected, respectively. In 47.8 % of cases, EGFR up-regulation and PTEN down-regulation occurred together. Survival was significantly poorer in tumors lacking GHRH expression. This worse prognosis in GHRH negative group remained significant even if SV1 was also expressed.

Conclusion

Our study shows that GHRH and SV1 genes expressed in human GB samples and their expression patterns are associated with poorer prognosis.
Literature
go back to reference Ang C, Guiot MC, Ramanakumar AV, Roberge D, Kavan P (2010) Clinical significance of molecular biomarkers in glioblastoma. Can J Neurol Sci 37(5):625–630PubMed Ang C, Guiot MC, Ramanakumar AV, Roberge D, Kavan P (2010) Clinical significance of molecular biomarkers in glioblastoma. Can J Neurol Sci 37(5):625–630PubMed
go back to reference Busto R, Schally AV, Braczkowski R, Plonowski A, Krupa M, Groot K, Armatis P, Varga JL (2002a) Expression of mRNA for growth hormone-releasing hormone and splice variants of GHRH receptors in human malignant bone tumors. Regul Pept 108:47–53PubMedCrossRef Busto R, Schally AV, Braczkowski R, Plonowski A, Krupa M, Groot K, Armatis P, Varga JL (2002a) Expression of mRNA for growth hormone-releasing hormone and splice variants of GHRH receptors in human malignant bone tumors. Regul Pept 108:47–53PubMedCrossRef
go back to reference Busto R, Schally AV, Varga JL, Garcia-Fernandez MO, Groot K, Armatis P, Szepeshazi K (2002b) The expression of growth hormone-releasing hormone (GHRH) and splice variants of its receptor in human gastroenteropancreatic carcinomas. Proc Natl Acad Sci USA 99:11866–11871PubMedCrossRefPubMedCentral Busto R, Schally AV, Varga JL, Garcia-Fernandez MO, Groot K, Armatis P, Szepeshazi K (2002b) The expression of growth hormone-releasing hormone (GHRH) and splice variants of its receptor in human gastroenteropancreatic carcinomas. Proc Natl Acad Sci USA 99:11866–11871PubMedCrossRefPubMedCentral
go back to reference Cancer Genome Atlas Research Network (2008) Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 455(7216):1061–1068CrossRef Cancer Genome Atlas Research Network (2008) Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 455(7216):1061–1068CrossRef
go back to reference Farkas R, Pozsgai E, Schally AV, Szigeti A, Szigeti E, Laszlo Z, Papp A, Gomori E, Mangel L, Horvath PO et al (2012) Possible predictors of histopathological response to neoadjuvant chemoradiotherapy for rectal cancer. J Cancer Res Clin Oncol 138(3):387–395PubMedCrossRef Farkas R, Pozsgai E, Schally AV, Szigeti A, Szigeti E, Laszlo Z, Papp A, Gomori E, Mangel L, Horvath PO et al (2012) Possible predictors of histopathological response to neoadjuvant chemoradiotherapy for rectal cancer. J Cancer Res Clin Oncol 138(3):387–395PubMedCrossRef
go back to reference Granata R, Trovato L, Gallo MP, Destefanis S, Settanni F, Scarlatti F, Brero A, Ramella R, Volante M, Isgaard J et al (2009) Growth hormone-releasing hormone promotes survival of cardiac myocytes in vitro and protects against ischaemia-reperfusion injury in rat heart. Cardiovasc Res 83(2):303–312PubMedCrossRef Granata R, Trovato L, Gallo MP, Destefanis S, Settanni F, Scarlatti F, Brero A, Ramella R, Volante M, Isgaard J et al (2009) Growth hormone-releasing hormone promotes survival of cardiac myocytes in vitro and protects against ischaemia-reperfusion injury in rat heart. Cardiovasc Res 83(2):303–312PubMedCrossRef
go back to reference Halmos G, Schally AV, Varga JL, Plonowski A, Rekasi Z, Czompoly T (2000) Human renal cell carcinoma expresses distinct binding sites for growth hormone-releasing hormone. Proc Natl Acad Sci USA 97(19):10555–10560PubMedCrossRefPubMedCentral Halmos G, Schally AV, Varga JL, Plonowski A, Rekasi Z, Czompoly T (2000) Human renal cell carcinoma expresses distinct binding sites for growth hormone-releasing hormone. Proc Natl Acad Sci USA 97(19):10555–10560PubMedCrossRefPubMedCentral
go back to reference Halmos G, Schally AV, Czompoly T, Krupa M, Varga JL, Rekasi Z (2002) Expression of growth hormone-releasing hormone and its receptor splice variants in human prostate cancer. J Clin Endocrinol Metab 87(10):4707–4714PubMedCrossRef Halmos G, Schally AV, Czompoly T, Krupa M, Varga JL, Rekasi Z (2002) Expression of growth hormone-releasing hormone and its receptor splice variants in human prostate cancer. J Clin Endocrinol Metab 87(10):4707–4714PubMedCrossRef
go back to reference Havt A, Schally AV, Halmos G, Varga JL, Toller GL, Horvath JE, Szepeshazi K, Köster F, Kovitz K, Groot K et al (2005) The expression of the pituitary growth hormone-releasing hormone receptor and its splice variants in normal and neoplastic human tissues. Proc Natl Acad Sci USA 102(48):17424–17429PubMedCrossRefPubMedCentral Havt A, Schally AV, Halmos G, Varga JL, Toller GL, Horvath JE, Szepeshazi K, Köster F, Kovitz K, Groot K et al (2005) The expression of the pituitary growth hormone-releasing hormone receptor and its splice variants in normal and neoplastic human tissues. Proc Natl Acad Sci USA 102(48):17424–17429PubMedCrossRefPubMedCentral
go back to reference 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 USA 102(35):12495–12500PubMedCrossRefPubMedCentral 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 USA 102(35):12495–12500PubMedCrossRefPubMedCentral
go back to reference Jaszberenyi M, Schally AV, Block NL, Zarandi M, Cai RZ, Vidaurre I, Szalontay L, Jayakumar AR, Rick FG (2013) Suppression of the proliferation of human U-87 MG glioblastoma cells by new antagonists of growth hormone. Releasing hormone in vivo and in vitro. Target Oncol 8(4):281–290. doi:10.1007/s11523-013-0264-y PubMedCrossRef Jaszberenyi M, Schally AV, Block NL, Zarandi M, Cai RZ, Vidaurre I, Szalontay L, Jayakumar AR, Rick FG (2013) Suppression of the proliferation of human U-87 MG glioblastoma cells by new antagonists of growth hormone. Releasing hormone in vivo and in vitro. Target Oncol 8(4):281–290. doi:10.​1007/​s11523-013-0264-y PubMedCrossRef
go back to reference 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):242–250PubMedCrossRefPubMedCentral 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):242–250PubMedCrossRefPubMedCentral
go back to reference Kita D, Yonekawa Y, Weller M, Ohgaki H (2007) PI3KCA alterations in primary (de novo) and secondary glioblastomas. Acta Neuropathol Berlin 113:295–302CrossRef Kita D, Yonekawa Y, Weller M, Ohgaki H (2007) PI3KCA alterations in primary (de novo) and secondary glioblastomas. Acta Neuropathol Berlin 113:295–302CrossRef
go back to reference Knobbe CB, Merlo A, Reifenberger G (2002) PTEN signaling in gliomas. Neuro Oncol J 4(3):196–211CrossRef Knobbe CB, Merlo A, Reifenberger G (2002) PTEN signaling in gliomas. Neuro Oncol J 4(3):196–211CrossRef
go back to reference Kreisl TN, Lassman AB, Mischel PS, Rosen N, Scher HI, Teruya-Feldstein J, Shaffer D, Lis E, Abrey LE (2009) A pilot study of everolimus and gefitinib in the treatment of recurrent glioblastoma (GBM). J Neurooncol 92(1):99–105PubMedCrossRef Kreisl TN, Lassman AB, Mischel PS, Rosen N, Scher HI, Teruya-Feldstein J, Shaffer D, Lis E, Abrey LE (2009) A pilot study of everolimus and gefitinib in the treatment of recurrent glioblastoma (GBM). J Neurooncol 92(1):99–105PubMedCrossRef
go back to reference Li J, Yen C, Liaw D, Podsypanina K, Bose S, Wang SI, Puc J, Miliaresis C, Rodgers L, McCombie R et al (1997) PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer. Science 275(5308):1943–1947PubMedCrossRef Li J, Yen C, Liaw D, Podsypanina K, Bose S, Wang SI, Puc J, Miliaresis C, Rodgers L, McCombie R et al (1997) PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer. Science 275(5308):1943–1947PubMedCrossRef
go back to reference Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25(4):402–408PubMedCrossRef Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25(4):402–408PubMedCrossRef
go back to reference Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW, Kleihues P (2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathologica 114(2):97–109. Erratum in: Acta Neuropathologica 114(5):547 Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW, Kleihues P (2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathologica 114(2):97–109. Erratum in: Acta Neuropathologica 114(5):547
go back to reference Maruno K, Absood A, Said SI (1998) Vasoactive intestinal peptide inhibits human small-cell lung cancer proliferation in vitro and in vivo. Proc Natl Acad Sci USA24;95(24):14373–14378 Maruno K, Absood A, Said SI (1998) Vasoactive intestinal peptide inhibits human small-cell lung cancer proliferation in vitro and in vivo. Proc Natl Acad Sci USA24;95(24):14373–14378
go back to reference Mellinghoff IK, Wang MY, Vivanco I, Haas-Kogan DA, Zhu S, Dia EQ et al (2005) Molecular determinants of the response of glioblastomas to EGFR kinase inhibitors. N Engl J Med 353:2012–2024PubMedCrossRef Mellinghoff IK, Wang MY, Vivanco I, Haas-Kogan DA, Zhu S, Dia EQ et al (2005) Molecular determinants of the response of glioblastomas to EGFR kinase inhibitors. N Engl J Med 353:2012–2024PubMedCrossRef
go back to reference Ohgaki H, Dessen P, Jourde B, Horstmann S, Nishikawa T, Di Patre PL, Burkhard C, Schüler D, Probst-Hensch NM, Maiorka PC (2004) Genetic pathways to glioblastoma: a population-based study. Cancer Res 64(19):6892–6899PubMedCrossRef Ohgaki H, Dessen P, Jourde B, Horstmann S, Nishikawa T, Di Patre PL, Burkhard C, Schüler D, Probst-Hensch NM, Maiorka PC (2004) Genetic pathways to glioblastoma: a population-based study. Cancer Res 64(19):6892–6899PubMedCrossRef
go back to reference Øvstebø R, Haug KB, Lande K, Kierulf P (2003) PCR-based Calibration Curves for Studies of Quantitative Gene Expression in Human Monocytes: development and Evaluation. Clin Chem 49(3):425–432PubMedCrossRef Øvstebø R, Haug KB, Lande K, Kierulf P (2003) PCR-based Calibration Curves for Studies of Quantitative Gene Expression in Human Monocytes: development and Evaluation. Clin Chem 49(3):425–432PubMedCrossRef
go back to reference 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):2313–2322PubMedCrossRef 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):2313–2322PubMedCrossRef
go back to reference Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. In: Misener S, Totowa NJ (eds) Krawetz S. Methods in Molecular Biology, Humana Press, Bioinformatics Methods and Protocols, pp 365–386 Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. In: Misener S, Totowa NJ (eds) Krawetz S. Methods in Molecular Biology, Humana Press, Bioinformatics Methods and Protocols, pp 365–386
go back to reference Schally AV (2008) New approaches to the therapy of various tumors based on peptide analogues. Horm Metab Res 40:315–322PubMedCrossRef Schally AV (2008) New approaches to the therapy of various tumors based on peptide analogues. Horm Metab Res 40:315–322PubMedCrossRef
go back to reference Schally AV, Halmos G (2012) Targeting to Peptide Receptors. In: Kratz F, Senter P, Steinhagen H (eds) Drug Delivery in Oncology: From Basic Research to Cancer Therapy. Wiley-VCH, Weinheim, pp 1219–1262 Schally AV, Halmos G (2012) Targeting to Peptide Receptors. In: Kratz F, Senter P, Steinhagen H (eds) Drug Delivery in Oncology: From Basic Research to Cancer Therapy. Wiley-VCH, Weinheim, pp 1219–1262
go back to reference Schally AV, Varga JL (1999) Antagonistic analogs of growth hormone-releasing hormone: new potential antitumor agents. Trends Endocrinol Metab 10:383–391PubMedCrossRef Schally AV, Varga JL (1999) Antagonistic analogs of growth hormone-releasing hormone: new potential antitumor agents. Trends Endocrinol Metab 10:383–391PubMedCrossRef
go back to reference Schally AV, Varga JL (2006) Antagonists of growth hormone-releasing hormone in oncology. Comb Chem High Throughput Screening 9(3):163–170CrossRef Schally AV, Varga JL (2006) Antagonists of growth hormone-releasing hormone in oncology. Comb Chem High Throughput Screening 9(3):163–170CrossRef
go back to reference Schally AV, Varga JL, Engel JB (2008) Antagonists of growth hormone-releasing hormone: an emerging new therapy for cancer. Nat Clin Pract Endocrinol Metab 4:33–43PubMedCrossRef Schally AV, Varga JL, Engel JB (2008) Antagonists of growth hormone-releasing hormone: an emerging new therapy for cancer. Nat Clin Pract Endocrinol Metab 4:33–43PubMedCrossRef
go back to reference Seitz S, Hohla F, Schally AV, Moder A, Engel JB, Horn F, Varga J, Zarandi M, Ortmann O, Köster F, Buchholz S (2008) Inhibition of estrogen receptor positive and negative breast cancer cell lines with a growth hormone-releasing hormone antagonist. Oncol Rep 20(5):1289–1294PubMed Seitz S, Hohla F, Schally AV, Moder A, Engel JB, Horn F, Varga J, Zarandi M, Ortmann O, Köster F, Buchholz S (2008) Inhibition of estrogen receptor positive and negative breast cancer cell lines with a growth hormone-releasing hormone antagonist. Oncol Rep 20(5):1289–1294PubMed
go back to reference Steck PA, Pershouse MA, Jasser SA, Yung WK, Lin H, Ligon AH, Langford LA, Baumgard ML, Hattier T, Davis T et al (1997) Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers. Nat Genet 15(4):356–362PubMedCrossRef Steck PA, Pershouse MA, Jasser SA, Yung WK, Lin H, Ligon AH, Langford LA, Baumgard ML, Hattier T, Davis T et al (1997) Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers. Nat Genet 15(4):356–362PubMedCrossRef
go back to reference Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U et al (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. European Organisation for Research and Treatment of Cancer Brain Tumor and Radiotherapy Groups; National Cancer Institute of Canada Clinical Trials Group. N Engl J Med 352(10):987–996PubMedCrossRef Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U et al (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. European Organisation for Research and Treatment of Cancer Brain Tumor and Radiotherapy Groups; National Cancer Institute of Canada Clinical Trials Group. N Engl J Med 352(10):987–996PubMedCrossRef
go back to reference Szepeshazi K, Schally AV, Groot K, Armatis P, Hebert F, Halmos G (2000) Antagonists of growth hormone-releasing hormone (GH-RH) inhibit in vivo proliferation of experimental pancreatic cancers and decrease IGF-II levels in tumours. Eur J Cancer 36:128–136PubMedCrossRef Szepeshazi K, Schally AV, Groot K, Armatis P, Hebert F, Halmos G (2000) Antagonists of growth hormone-releasing hormone (GH-RH) inhibit in vivo proliferation of experimental pancreatic cancers and decrease IGF-II levels in tumours. Eur J Cancer 36:128–136PubMedCrossRef
go back to reference Vacas E, Fernández-Martínez AB, Bajo AM, Sánchez-Chapado M, Schally AV, Prieto JC, Carmena MJ (2012) Vasoactive intestinal peptide (VIP) inhibits human renal cell carcinoma proliferation. Biochim Biophys Acta 1823(10):1676–1685PubMedCrossRef Vacas E, Fernández-Martínez AB, Bajo AM, Sánchez-Chapado M, Schally AV, Prieto JC, Carmena MJ (2012) Vasoactive intestinal peptide (VIP) inhibits human renal cell carcinoma proliferation. Biochim Biophys Acta 1823(10):1676–1685PubMedCrossRef
go back to reference Weller M, Felsberg J, Hartmann C, Berger H, Steinbach JP, Schramm J, Westphal M, Schackert G, Simon M, Tonn JC, Heese O, Krex D, Nikkhah G, Pietsch T, Wiestler O, Reifenberger G, von Deimling A, Loeffler M (2009) Molecular predictors of progression-free and overall survival in patients with newly diagnosed glioblastoma: a prospective translational study of the German Glioma Network. J Clin Oncol 27(34):5743–5750PubMedCrossRef Weller M, Felsberg J, Hartmann C, Berger H, Steinbach JP, Schramm J, Westphal M, Schackert G, Simon M, Tonn JC, Heese O, Krex D, Nikkhah G, Pietsch T, Wiestler O, Reifenberger G, von Deimling A, Loeffler M (2009) Molecular predictors of progression-free and overall survival in patients with newly diagnosed glioblastoma: a prospective translational study of the German Glioma Network. J Clin Oncol 27(34):5743–5750PubMedCrossRef
go back to reference Zhu H, Acquaviva J, Ramachandran P, Boskovitz A, Woolfenden S, Pfannl R, Bronson RT, Chen JW, Weissleder R, Housman DE et al (2009) Oncogenic EGFR signaling cooperates with loss of tumor suppressor gene functions in gliomagenesis. Proc Natl Acad Sci USA 106(8):2712–2716PubMedCrossRefPubMedCentral Zhu H, Acquaviva J, Ramachandran P, Boskovitz A, Woolfenden S, Pfannl R, Bronson RT, Chen JW, Weissleder R, Housman DE et al (2009) Oncogenic EGFR signaling cooperates with loss of tumor suppressor gene functions in gliomagenesis. Proc Natl Acad Sci USA 106(8):2712–2716PubMedCrossRefPubMedCentral
Metadata
Title
Prognosis in human glioblastoma based on expression of ligand growth hormone-releasing hormone, pituitary-type growth hormone-releasing hormone receptor, its splicing variant receptors, EGF receptor and PTEN genes
Authors
Géza Mezey
Andrea Treszl
Andrew V. Schally
Normann L. Block
Laura Vízkeleti
Alíz Juhász
Álmos Klekner
János Nagy
Margit Balázs
Gábor Halmos
László Bognár
Publication date
01-10-2014
Publisher
Springer Berlin Heidelberg
Published in
Journal of Cancer Research and Clinical Oncology / Issue 10/2014
Print ISSN: 0171-5216
Electronic ISSN: 1432-1335
DOI
https://doi.org/10.1007/s00432-014-1716-1

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