The Effect of LHRH Antagonist Cetrorelix in Crossover Conditioned Media from Epithelial (BPH-1) and Stromal (WPMY-1) Prostate Cells

Abstract

Stromal cells strictly modulate the differentiation of the normal prostate epithelium. In benign prostatic hyperplasia (BPH) tissue, the ratio of stromal to epithelial cells reaches a 5:1 ratio. In this study, we evaluated the effects of crossover conditioned media (CM) of stromal and epithelial prostate cells before and after treatment with LHRH antagonist Cetrorelix. WPMY-1 human prostate stromal cells and BPH-1 human benign prostatic hyperplasia cells were cultured in vitro and the effects of crossover conditioned media (CM) from those cells were studied. We evaluated the effect of Cetrorelix on the expression of PCNA and p53 in those cells. We then studied the effect of Cetrorelix on BPH-1 cells cultured with the CM from WPMY-1 cells, as well as the mechanisms which govern these interactions. CM from WPMY-1 cells strongly stimulated the proliferation of BPH-1 cells in a dose dependent manner, while CM from BPH-1 cells only slightly increased the proliferation of WPMY-1 cells. Cetrorelix inhibited the proliferation of both cell lines and the expression of PCNA, while the expression of p53 was increased. Cetrorelix also inhibited the proliferation of BPH-1 cells stimulated with the CM from WPMY-1 cells. In the crossover experiment, conditioned media from WPMY-1 and BPH-1 cells increased the expression of phosphorylated ERK1/2 and STAT3. Our results support previous observations on the bidirectional stromal-epithelial interactions in prostate gland and shed more light on the mechanistic action of those effects. Our study strongly supports the hypothesis that LHRH antagonists may be beneficial for BPH prevention and treatment.

Key words

LHRH receptor - LHRH antagonist - Cetrorelix - benign prostatic hyperplasia - WPMY-1 - BPH-1

• References

• 1 Price H, McNeal JE, Stamey TA. Evolving patterns of tissue composition in benign prostatic hyperplasia as a function of specimen size. Hum Pathol 1990; 21: 578-585
• 2 Starsichova A, Lincova E, Pernicova Z, Kozubik A, Soucek K. TGF-beta1 suppresses IL-6-induced STAT3 activation through regulation of Jak2 expression in prostate epithelial cells. Cell Signal 2010; 22: 1734-1744
• 3 Schauer IG, Rowley DR. The functional role of reactive stroma in benign prostatic hyperplasia. Differentiation 2011; 82: 200-210
• 4 Barabutis N, Catravas JD. Anti-Inflammatory Activity of Hsp90 Inhibitors in the Human Vasculature. Med Surg Urol 2013; 2: e104
• 5 Alonso-Magdalena P, Brossner C, Reiner A, Cheng G, Sugiyama N, Warner M, Gustafsson JA. A role for epithelial-mesenchymal transition in the etiology of benign prostatic hyperplasia. Proc Natl Acad Sci USA 2009; 106: 2859-2863
• 6 Kapoor A. Benign prostatic hyperplasia (BPH) management in the primary care setting. Can J Urol 2012; 19: 10-17
• 7 Schally AV. New approaches to the therapy of various tumors based on peptide analogues. Horm Metab Res 2008; 40: 315-322
• 8 Siejka A, Schally AV, Block NL, Barabutis N. Mechanisms of inhibition of human benign prostatic hyperplasia in vitro by the luteinizing hormone-releasing hormone antagonist cetrorelix. BJU Int 2010; 106: 1382-1388
• 9 Li W, Wu CL, Febbo PG, Olumi AF. Stromally expressed c-Jun regulates proliferation of prostate epithelial cells. Am J Pathol 2007; 171: 1189-1198
• 10 Webber MM, Trakul N, Thraves PS, Bello-DeOcampo D, Chu WW, Storto PD, Huard TK, Rhim JS, Williams DE. A human prostatic stromal myofibroblast cell line WPMY-1: a model for stromal-epithelial interactions in prostatic neoplasia. Carcinogenesis 1999; 20: 1185-1192
• 11 Palapattu GS, Sutcliffe S, Bastian PJ, Platz EA, De Marzo AM, Isaacs WB, Nelson WG. Prostate carcinogenesis and inflammation: emerging insights. Carcinogenesis 2005; 26: 1170-1181
• 12 De Marzo AM, Platz EA, Sutcliffe S, Xu J, Gronberg H, Drake CG, Nakai Y, Isaacs WB, Nelson WG. Inflammation in prostate carcinogenesis. Nat Rev Cancer 2007; 7: 256-269
• 13 Hayward SW, Dahiya R, Cunha GR, Bartek J, Deshpande N, Narayan P. Establishment and characterization of an immortalized but non-transformed human prostate epithelial cell line: BPH-1. In Vitro Cell Dev Biol Anim 1995; 31: 14-24
• 14 Bajusz S, Csernus VJ, Janaky T, Bokser L, Fekete M, Schally AV. New antagonists of LHRH. II. Inhibition and potentiation of LHRH by closely related analogues. Int J Pept Protein Res 1988; 32: 425-435
• 15 Barabutis N, Schally AV. Knocking down gene expression for growth hormone-releasing hormone inhibits proliferation of human cancer cell lines. Br J Cancer 2008; 98: 1790-1796
• 16 Barabutis N, Siejka A, Schally AV, Block NL, Cai R, Varga JL. Activation of mitogen-activated protein kinases by a splice variant of GHRH receptor. J Mol Endocrinol 2010; 44: 127-134
• 17 Siejka A, Lawnicka H, Komorowski J, Schally AV, Stepień T, Krupiński R, Stepień H. GH-RH antagonist (MZ-4-71) inhibits VEGF secretion and proliferation of murine endothelial cells. Life Sci 2003; 72: 2473-2479
• 18 Lepor H. Pathophysiology, epidemiology, and natural history of benign prostatic hyperplasia. Rev Urol 2004; 6: S3-S10
• 19 Roehrborn CG. BPH progression: concept and key learning from MTOPS, ALTESS, COMBAT, and ALF-ONE. BJU Int 2008; 101: 17-21
• 20 Auffenberg GB, Helfand BT, McVary KT. Established medical therapy for benign prostatic hyperplasia. Urol Clin North Am 2009; 36: 443-459 v–vi
• 21 Siejka A, Schally AV, Block NL, Barabutis N. Antagonists of growth hormone-releasing hormone inhibit the proliferation of human benign prostatic hyperplasia cells. Prostate 2010; 70: 1087-1093
• 22 Mebratu Y, Tesfaigzi Y. How ERK1/2 activation controls cell proliferation and cell death: Is subcellular localization the answer?. Cell Cycle 2009; 8: 1168-1175
• 23 Kyriakis JM, Avruch J. Mammalian MAPK signal transduction pathways activated by stress and inflammation: a 10-year update. Physiol Rev 2012; 92: 689-737
• 24 Yu H, Pardoll D, Jove R. STATs in cancer inflammation and immunity: a leading role for STAT3. Nat Rev Cancer 2009; 9: 798-809
• 25 Barabutis N, Schally AV. Growth hormone-releasing hormone: extrapituitary effects in physiology and pathology. Cell Cycle 2010; 9: 4110-4116
• 26 Huang HF, Murphy TF, Shu P, Barton AB, Barton BE. Stable expression of constitutively-activated STAT3 in benign prostatic epithelial cells changes their phenotype to that resembling malignant cells. Mol Cancer 2005; 4: 2
• 27 Moldovan GL, Pfander B, Jentsch S. PCNA, the maestro of the replication fork. Cell 2007; 129: 665-679
• 28 Wang W, Bergh A, Damber JE. Chronic inflammation in benign prostate hyperplasia is associated with focal upregulation of cyclooxygenase-2, Bcl-2, and cell proliferation in the glandular epithelium. Prostate 2004; 61: 60-72
• 29 Shimizu I, Yoshida Y, Katsuno T, Tateno K, Okada S, Moriya J, Yokoyama M, Nojima A, Ito T, Zechner R, Komuro I, Kobayashi Y, Minamino T. p53-induced adipose tissue inflammation is critically involved in the development of insulin resistance in heart failure. Cell Metab 2012; 15: 51-64
• 30 Schlechte H, Lenk SV, Loning T, Schnorr D, Rudolph BD, Ditscherlein G, Loening SA. p53 tumour suppressor gene mutations in benign prostatic hyperplasia and prostate cancer. Eur Urol 1998; 34: 433-440
• 31 Addadi Y, Moskovits N, Granot D, Lozano G, Carmi Y, Apte RN, Neeman M, Oren M. p53 status in stromal fibroblasts modulates tumor growth in an SDF1-dependent manner. Cancer Res 2010; 70: 9650-9658
• 32 Hill R, Song Y, Cardiff RD, Van Dyke T. Selective evolution of stromal mesenchyme with p53 loss in response to epithelial tumorigenesis. Cell 2005; 123: 1001-1011
• 33 Trimis G, Chatzistamou I, Politi K, Kiaris H, Papavassiliou AG. Expression of p21waf1/Cip1 in stromal fibroblasts of primary breast tumors. Hum Mol Genet 2008; 17: 3596-3600
• 34 Rick FG, Schally AV, Block NL, Abi-Chaker A, Krishan A, Szalontay L. Mechanisms of synergism between antagonists of growth hormone-releasing hormone and antagonists of luteinizing hormone-releasing hormone in shrinking experimental benign prostatic hyperplasia. Prostate 2012; 73: 873-883
• 35 Rick FG, Schally AV, Block NL, Halmos G, Perez R, Fernandez JB, Vidaurre I, Szalontay L. LHRH antagonist Cetrorelix reduces prostate size and gene expression of proinflammatory cytokines and growth factors in a rat model of benign prostatic hyperplasia. Prostate 2011; 71: 736-747
• 36 Rick FG, Abi-Chaker A, Szalontay L, Perez R, Jaszberenyi M, Jayakumar AR, Shamaladevi N, Szepeshazi K, Vidaurre I, Halmos G, Krishan A, Block NL, Schally AV. Shrinkage of experimental benign prostatic hyperplasia and reduction of prostatic cell volume by a gastrin-releasing peptide antagonist. Proc Natl Acad Sci USA 2013; 110: 2617-2622
• 37 Petronilho F, Vuolo F, Galant LS, Constantino L, Tomasi CD, Giombelli VR, de Souza CT, da silva S, Barbeiro DF, Soriano FG, Streck EL, Ritter C, Zanotto-Filho A, Pasquali MA, Gelain DP, Rybarczyk-Filho JL, Moreira JC, Block NL, Roesler R, Schwartsmann G, Schally AV, Dal-Pizzol F. Gastrin-releasing peptide receptor antagonism induces protection from lethal sepsis: involvement of toll-like receptor 4 signaling. Mol Med 2012; 18: 1209-1219
• 38 Lucas R, Sridhar S, Rick FG, Gorshkov B, Umapathy NS, Yang G, Oseghale A, Verin AD, Chakraborty T, Matthay MA, Zemskov EA, White R, Block NL, Schally AV. Agonist of growth hormone-releasing hormone reduces pneumolysin-induced pulmonary permeability edema. Proc Natl Acad Sci USA 2012; 109: 2084-2089
• 39 Barabutis N, Schally AV. Antioxidant activity of growth hormone-releasing hormone antagonists in LNCaP human prostate cancer line. Proc Natl Acad Sci USA 2008; 105: 20470-20475
• 40 Barabutis N, Siejka A, Schally AV. Growth hormone releasing hormone induces the expression of nitric oxide synthase. J Cell Mol Med 2011; 15: 1148-1155
• 41 Rick FG, Szalontay L, Schally AV, Block NL, Nadji M, Szepeshazi K, Vidaurre I, Zarandi M, Kovacs M, Rekasi Z. Combining growth hormone-releasing hormone antagonist with luteinizing hormone-releasing hormone antagonist greatly augments benign prostatic hyperplasia shrinkage. J Urol 2012; 187: 1498-1504