Inhibition of growth hormone receptor by Somavert reduces expression of GPER and prevents growth stimulation of triple‑negative breast cancer by 17β‑estradiol

Girgert,Rainer; Emons,Günter; Gründker,Carsten

doi:10.3892/ol.2018.8521

Inhibition of growth hormone receptor by Somavert reduces expression of GPER and prevents growth stimulation of triple‑negative breast cancer by 17β‑estradiol

Authors:
- Rainer Girgert
- Günter Emons
- Carsten Gründker
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, University Medical Center Goettingen, D‑37075 Goettingen, Germany
Published online on: April 18, 2018 https://doi.org/10.3892/ol.2018.8521
Pages: 9559-9566

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Currently, conventional chemotherapy is the only treatment option for triple‑negative breast cancers (TNBC) due to a lack of a unique target. In TNBC, a high expression of the membrane bound G protein‑coupled estrogen receptor (GPER), correlates with a worse outcome. There is a potential for an association between growth hormone receptor (GHR) and GPER expression. To confirm this hypothesis, GHR was inhibited in TNBC cells with Somavert, and GPER expression levels, and the effect on signal transduction and proliferation induction in TNBC cells were analyzed. Proliferation of TNBC cells was measured using an Alamar‑blue assay. Expression of GPER and activation of c‑src and epidermal growth factor receptor (EGFR) by 17β‑estradiol was analyzed by western blotting. Induction of c‑fos, cyclin D1 and aromatase expression was determined by reverse transcription‑semi‑quantitative polymerase chain reaction. The expression of GPER was concentration‑ and time‑dependently reduced by Somavert down to 46±7% (P<0.01) of the control. Furthermore, 17β‑estradiol significantly increased the cell number of HCC1806 cells to 128±14% (P<0.05), and that of MDA‑MB‑453 cells to 115±3%. This increase in cell number was reduced to 103±11% in HCC1806 cells in which GPER expression was downregulated by Somavert, and to 102±3% in MDA‑MB‑453 cells. In addition, 17β‑estradiol increased the activation of c‑src in HCC1806 cells by 1.8‑fold, and Somavert reduced p‑src to 63% of control. In MDA‑MB‑453 cells src phosphorylation increased by 7‑fold upon stimulation with estradiol, but after treatment with Somavert only a 4‑fold increase was observed. Phosphorylation of EGFR was increased by 2.2‑fold of control in HCC1806 cells by 17β‑estradiol, and by 1.4‑fold in MDA‑MD‑453 cells. Somavert completely prevented this activation. Induction of cyclin D1 and aromatase expression by 17β‑estradiol was also prevented by Somavert. Somavert reduces GPER expression in triple negative breast cancer cells. Treatment with Somavert prevents induction of genes regulating proliferation by 17β‑estradiol. Inhibition of GPER expression is a promising therapeutic intervention for TNBC.

View Figures

View References

1	Dowsett M, Cuzick J, Ingle J, Coates A, Forbes J, Bliss J, Buyse M, Baum M, Buzdar A, Colleoni M, et al: Meta-analysis of breast cancer outcomes in adjuvant trials of aromatase inhibitors versus tamoxifen. J Clin Oncol. 28:509–518. 2010. View Article : Google Scholar : PubMed/NCBI
2	Carey LA, Dees EC, Sawyer L, Gatti L, Moore DT, Collichio F, Ollila DW, Sartor CI, Graham ML and Perou CM: The triple negative paradox: Primary tumor chemosensitivity of breast cancer subtypes. Clin Cancer Res. 13:2329–2334. 2007. View Article : Google Scholar : PubMed/NCBI
3	Silver DP, Richardson AL, Eklund AC, Wang ZC, Szallasi Z, Li Q, Juul N, Leong CO, Calogrias D, Buraimoh A, et al: Efficacy of neoadjuvant Cisplatin in triple-negative breast cancer. J Clin Oncol. 28:1145–1153. 2010. View Article : Google Scholar : PubMed/NCBI
4	Reis-Filho JS and Tutt AN: Triple negative tumours: A critical review. Histopathology. 52:108–118. 2008. View Article : Google Scholar : PubMed/NCBI
5	Baselga J, Gómez P, Greil R, Braga S, Climent MA, Wardley AM, Kaufman B, Stemmer SM, Pego A, Chan A, et al: Randomized phase II study of the anti-epidermal growth factor receptor monoclonal antibody cetuximab with cisplatin versus cisplatin alone in patients with metastatic triple-negative breast cancer. J Clin Oncol. 31:2586–2592. 2013. View Article : Google Scholar : PubMed/NCBI
6	Scaling AL, Prossnitz ER and Hathaway HJ: GPER mediates estrogen-induced signaling and proliferation in human breast epithelial cells and normal and malignant breast. Horm Cancer. 5:146–160. 2014. View Article : Google Scholar : PubMed/NCBI
7	Steiman J, Peralta EA, Louis S and Kamel O: Biology of the estrogen receptor, GPR30, in triple negative breast cancer. Am J Surg. 206:698–703. 2013. View Article : Google Scholar : PubMed/NCBI
8	Girgert R, Emons G and Gründker C: Inactivation of GPR30 reduces growth of triple-negative breast cancer cells: Possible application in targeted therapy. Breast Cancer Res Treat. 134:199–205. 2012. View Article : Google Scholar : PubMed/NCBI
9	Girgert R, Emons G and Gründker C: Inhibition of GPR30 by estriol prevents growth stimulation of triple-negative breast cancer cells by 17β-estradiol. BMC Cancer. 14:9352014. View Article : Google Scholar : PubMed/NCBI
10	Vivacqua A, Lappano R, De Marco P, Sisci D, Aquila S, De Amicis F, Fuqua SA, Andó S and Maggiolini M: G protein-coupled receptor 30 expression is up-regulated by EGF and TGF alpha in estrogen receptor alpha-positive cancer cells. Mol Endocrinol. 23:1815–1826. 2009. View Article : Google Scholar : PubMed/NCBI
11	Nielsen TO, Hsu FD, Jensen K, Cheang M, Karaca G, Hu Z, Hernandez-Boussard T, Livasy C, Cowan D, Dressler L, et al: Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma. Clin Cancer Res. 10:5367–5374. 2004. View Article : Google Scholar : PubMed/NCBI
12	Girgert R, Emons G and Gründker C: 17β-estradiol-induced growth of triple-negative breast cancer cells is prevented by the reduction of GPER expression after treatment with gefitinib. Oncol Rep. 37:1212–1218. 2017. View Article : Google Scholar : PubMed/NCBI
13	Divisova J, Kuiatse I, Lazard Z, Weiss H, Vreeland F, Hadsell DL, Schiff R, Osborne CK and Lee AV: The growth hormone receptor antagonist pegvisomant blocks both mammary gland development and MCF-7 breast cancer xenograft growth. Breast Cancer Res Treat. 98:315–327. 2006. View Article : Google Scholar : PubMed/NCBI
14	Perez R, Schally AV, Vidaurre I, Rincon R, Block NL and Rick FG: Antagonists of growth hormone-releasing hormone suppress in vivo tumor growth and gene expression in triple negative breast cancers. Oncotarget. 3:988–997. 2012. View Article : Google Scholar : PubMed/NCBI
15	Neggers SJ, Franck SE, de Rooij FW, Dallenga AH, Poublon RM, Feelders RA, Janssen JA, Buchfelder M, Hofland LJ, Jorgensen JO, et al: Long-term efficacy and safety of pegvisomant in combination with long-acting somatostatin analogs in acromegaly. J Clin Endocrinol Metab. 99:3644–3652. 2014. View Article : Google Scholar : PubMed/NCBI
16	Neggers SJ and van der Lely AJ: Somatostatin analog and pegvisomant combination therapy for acromegaly. Nat Rev Endocrinol. 5:546–552. 2009. View Article : Google Scholar : PubMed/NCBI
17	Kireeva ML, Mo FE, Yang GP and Lau LF: Cyr61, a product of a growth factor-inducible immediate-early gene, promotes cell proliferation, migration, and adhesion. Mol Cell Biol. 16:1326–1334. 1996. View Article : Google Scholar : PubMed/NCBI
18	Girgert R, Bartsch C, Hill SM, Kreienberg R and Hanf V: Tracking the elusive antiestrogenic effect of melatonin: A new methodological approach. Neuro Endocrinol Lett. 24:440–444. 2003.PubMed/NCBI
19	Filardo EJ, Quinn JA, Frackelton AR Jr and Bland KI: Estrogen action via the G protein-coupled receptor, GPR30: Stimulation of adenylyl cyclase and cAMP-mediated attenuation of the epidermal growth factor receptor-to-MAPK signaling axis. Mol Endocrinol. 16:70–84. 2002. View Article : Google Scholar : PubMed/NCBI
20	Ignatov A, Ignatov T, Weissenborn C, Eggemann H, Bischoff J, Semczuk A, Roessner A, Costa SD and Kalinski T: G-protein-coupled estrogen receptor GPR30 and tamoxifen resistance in breast cancer. Breast Cancer Res Treat. 128:457–466. 2011. View Article : Google Scholar : PubMed/NCBI
21	Contreras B and Talamantes F: Growth hormone (GH) and 17beta-estradiol regulation of the expression of mouse GH receptor and GH-binding protein in cultured mouse hepatocytes. Endocrinology. 140:4725–4731. 1999. View Article : Google Scholar : PubMed/NCBI
22	Slootweg MC, Swolin D, Netelenbos JC, Isaksson OG and Ohlsson C: Estrogen enhances growth hormone receptor expression and growth hormone action in rat osteosarcoma cells and human osteoblast-like cells. J Endocrinol. 155:159–164. 1997. View Article : Google Scholar : PubMed/NCBI
23	Birzniece V, Sata A and Ho KK: Growth hormone receptor modulators. Rev Endocr Metab Disord. 10:145–156. 2009. View Article : Google Scholar : PubMed/NCBI