GPR30 expression and function in breast cancer cells are induced through a cis‑acting element targeted by ETS factors

Segura‑Bautista,David; Olivares,Aleida; Casas‑González,Patricia; Bonilla,Edmundo; Salazar,Zayil; Pérez‑Solis,Marco Allán

doi:10.3892/or.2020.7540

GPR30 expression and function in breast cancer cells are induced through a cis‑acting element targeted by ETS factors

Authors:
- David Segura‑Bautista
- Aleida Olivares
- Patricia Casas‑González
- Edmundo Bonilla
- Zayil Salazar
- Marco Allán Pérez‑Solis
View Affiliations

Affiliations: Medical Research Unit in Reproductive Medicine, UMAE Hospital de Gineco Obstetricia no. 4 ‘Luis Castelazo‑Ayala’, Instituto Mexicano del Seguro Social, México City 01090, México, Departament of Health Science, Universidad Autónoma Metropolitana, Iztapalapa, México City 09340, México, Biological and Health Sciences PhD Program, Universidad Autónoma Metropolitana, Iztapalapa, México City 09340, México
Published online on: March 10, 2020 https://doi.org/10.3892/or.2020.7540
Pages: 1669-1682

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

The capacity that G protein‑coupled receptor 30 (GPR30) has demonstrated for triggering estrogen‑dependent signaling pathways has attracted the interest of breast cancer researchers; however, the reported expression profiles and functions of GPR30 in breast cancer are inconsistent. The main purpose of the present investigation was to identify transcriptional mechanisms underlying the expression of GPR30 that allow a better understanding of its role in breast cancer progression. In the cell lines used as different polarity models in the present study, it was determined immunologically that GPR30 is expressed in normal mammary gland cells and that this expression decreased considerably during breast cancer development, where cell identity is lost. However, it was also determined that, in spite of low GPR30 expression levels in breast cancer cells with little differentiation, this membrane estrogen receptor (ER) is able to increase cell viability and suppress migration in cells that have acquired metastatic capacity. In addition, through transient expression assays in breast cancer cells, it was revealed that a transcriptional mechanism dependent on protein kinase A and susceptible to retinoic acid in ER‑positive cells induces GPR30 expression through a cis‑regulatory element for E26 transformation‑specific transcription factors, located between ‑631 and ‑625 bp from the GPR30 translation start codon. Overall, these results suggested that in vitro transcriptional regulation of GPR30 expression in breast cancer cells may serve a relevant role in the conservation of an epithelial phenotype, and also may be important to avoid the transition to metastasis.

View Figures

View References

1	Vergote I, Amant F, Leunen K, Van Gorp T, Berteloot P and Neven P: Metastatic breast cancer: Sequencing hormonal therapy and positioning of fulvestrant. Int J Gynecol Cancer. 16 (Suppl 2):S524–S526. 2006. View Article : Google Scholar
2	DeSantis CE, Ma J, Gaudet MM, Newman LA, Miller KD, Goding Sauer A, Jemal A and Siegel RL: Breast cancer statistics, 2019. CA Cancer J Clin. 69:438–451. 2019. View Article : Google Scholar : PubMed/NCBI
3	Fan W, Chang J and Fu P: Endocrine therapy resistance in breast cancer: Current status, possible mechanisms and overcoming strategies. Future Med Chem. 7:1511–1519. 2015. View Article : Google Scholar : PubMed/NCBI
4	Barton M, Filardo EJ, Lolait SJ, Thomas P, Maggiolini M and Prossnitz ER: Twenty years of the G protein-coupled estrogen receptor GPER: Historical and personal perspectives. J Steroid Biochem Mol Biol. 176:4–15. 2018. View Article : Google Scholar : PubMed/NCBI
5	Ruan SQ, Wang SW, Wang ZH and Zhang SZ: Regulation of HRG-β1-induced proliferation, migration and invasion of MCF-7 cells by upregulation of GPR30 expression. Mol Med Rep. 6:131–138. 2012.PubMed/NCBI
6	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
7	Molina L, Figueroa CD, Bhoola KD and Ehrenfeld P: GPER-1/GPR30 a novel estrogen receptor sited in the cell membrane: Therapeutic coupling to breast cancer. Expert Opin Ther Targets. 21:755–766. 2017. View Article : Google Scholar : PubMed/NCBI
8	Thomas P and Dong J: Binding and activation of the seven-transmembrane estrogen receptor GPR30 by environmental estrogens: A potential novel mechanism of endocrine disruption. J Steroid Biochem Mol Biol. 102:175–179. 2006. View Article : Google Scholar : PubMed/NCBI
9	Long N, Long B, Mana A, Le D, Nguyen L, Chokr S and Sinchak K: Tamoxifen and ICI 182,780 activate hypothalamic G protein-coupled estrogen receptor 1 to rapidly facilitate lordosis in female rats. Horm Behav. 89:98–103. 2017. View Article : Google Scholar : PubMed/NCBI
10	Bologa CG, Revankar CM, Young SM, Edwards BS, Arterburn JB, Kiselyov AS, Parker MA, Tkachenko SE, Savchuck NP, Sklar LA, et al: Virtual and biomolecular screening converge on a selective agonist for GPR30. Nat Chem Biol. 2:207–212. 2006. View Article : Google Scholar : PubMed/NCBI
11	Dennis MK, Burai R, Ramesh C, Petrie WK, Alcon SN, Nayak TK, Bologa CG, Leitao A, Brailoiu E, Deliu E, et al: In vivo effects of a GPR30 antagonist. Nat Chem Biol. 5:4212009. View Article : Google Scholar : PubMed/NCBI
12	Dennis MK, Field AS, Burai R, Ramesh C, Petrie WK, Bologa CG, Oprea TI, Yamaguchi Y, Hayashi S, Sklar LA, et al: Identification of a GPER/GPR30 antagonist with improved estrogen receptor counterselectivity. J Steroid Biochem Mol Biol. 127:358–366. 2011. View Article : Google Scholar : PubMed/NCBI
13	Mo Z, Liu M, Yang F, Luo H, Li Z, Tu G and Yang G: GPR30 as an initiator of tamoxifen resistance in hormone-dependent breast cancer. Breast Cancer Res. 15:R1142013. View Article : Google Scholar : PubMed/NCBI
14	Pandey DP, Lappano R, Albanito L, Madeo A, Maggiolini M and Picard D: Estrogenic GPR30 signalling induces proliferation and migration of breast cancer cells through CTGF. EMBO J. 28:523–532. 2009. View Article : Google Scholar : PubMed/NCBI
15	Lucki NC and Sewer MB: Genistein stimulates MCF-7 breast cancer cell growth by inducing acid ceramidase (ASAH1) gene expression. J Biol Chem. 86:19399–19409. 2011. View Article : Google Scholar
16	Recchia AG, De Francesco EM, Vivacqua A, Sisci D, Panno ML, Andò S and Maggiolini M: The G protein-coupled receptor 30 is up-regulated by hypoxia-inducible factor-1alpha (HIF-1alpha) in breast cancer cells and cardiomyocytes. J Biol Chem. 286:10773–10782. 2011. View Article : Google Scholar : PubMed/NCBI
17	De Francesco EM, Maggiolini M and Musti AM: Crosstalk between Notch, HIF-1α and GPER in Breast Cancer EMT. Int J Mol Sci. 19(pii): E20112018. View Article : Google Scholar : PubMed/NCBI
18	Ariazi EA, Brailoiu E, Yerrum S, Shupp HA, Slifker MJ, Cunliffe HE, Black MA, Donato AL, Arterburn JB, Oprea TI, et al: The G protein-coupled receptor GPR30 inhibits proliferation of estrogen receptor-positive breast cancer cells. Cancer Res. 70:1184–1194. 2010. View Article : Google Scholar : PubMed/NCBI
19	Chen ZJ, Wei W, Jiang GM, Liu H, Wei WD, Yang X, Wu YM, Liu H, Wong CK, Du J and Wang HS: Activation of GPER suppresses epithelial mesenchymal transition of triple negative breast cancer cells via NF-κB signals. Mol Oncol. 10:775–788. 2016. View Article : Google Scholar : PubMed/NCBI
20	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
21	Filigheddu N, Sampietro S, Chianale F, Porporato PE, Gaggianesi M, Gregnanin I, Rainero E, Ferrara M, Perego B, Riboni F, et al: Diacylglycerol kinase α mediates 17-β-estradiol-induced proliferation, motility, and anchorage-independent growth of Hec-1A endometrial cancer cell line through the G protein-coupled estrogen receptor GPR30. Cell Signal. 23:1988–1996. 2011. View Article : Google Scholar : PubMed/NCBI
22	Ge C, Yu M and Zhang C: G protein-coupled receptor 30 mediates estrogen-induced proliferation of primordial germ cells via EGFR/Akt/β-catenin signaling pathway. Endocrinology. 153:3504–3516. 2012. View Article : Google Scholar : PubMed/NCBI
23	Hsu LH, Chu NM, Lin YF and Kao SH: G-protein coupled estrogen receptor in breast cancer. Int J Mol Sci. 20(pii): E3062019. View Article : Google Scholar : PubMed/NCBI
24	Poola I, Abraham J, Liu A, Marshalleck JJ and DeWitty RL: The cell surface estrogen receptor, G protein-coupled receptor 30 (GPR30), is markedly down regulated during breast tumorigenesis. Breast Cancer (Auckl). 1:65–78. 2008.PubMed/NCBI
25	Wei W, Chen ZJ, Zhang KS, Yang XL, Wu YM, Chen XH, Huang HB, Liu HL, Cai SH, Du J and Wang HS: The activation of G protein-coupled receptor 30 (GPR30) inhibits proliferation of estrogen receptor-negative breast cancer cells in vitro and in vivo. Cell Death Dis. 5:e14282014. View Article : Google Scholar : PubMed/NCBI
26	Yu T, Liu M, Luo H, Wu C, Tang X, Tang S, Hu P, Yan Y, Wang Z and Tu G: GPER mediates enhanced cell viability and motility via non-genomic signaling induced by 17β-estradiol in triple-negative breast cancer cells. J Steroid Biochem Mol Biol. 143:392–403. 2014. View Article : Google Scholar : PubMed/NCBI
27	Schreiber E, Matthias P, Müller MM and Schaffner W: Rapid detection of octamer binding proteins with ‘mini-extracts’, prepared from a small number of cells. Nucleic Acids Res. 17:64191989. View Article : Google Scholar : PubMed/NCBI
28	Sambrook J, Fritsch EF and Maniatis T: Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press; pp. 1–3. 1989
29	Filardo EJ and Thomas P: Minireview: G protein-coupled estrogen receptor-1, GPER-1: Its mechanism of action and role in female reproductive cancer, renal and vascular physiology. Endocrinology. 153:2953–2962. 2012. View Article : Google Scholar : PubMed/NCBI
30	Janknecht R, Monté D, Baert JL and de Launoit Y: The ETS-related transcription factor ERM is a nuclear target of signaling cascades involving MAPK and PKA. Oncogene. 13:1745–1754. 1996.PubMed/NCBI
31	Wu J and Janknecht R: Regulation of the ETS transcription factor ER81 by the 90-kDa ribosomal S6 kinase 1 and protein kinase A. J Biol Chem. 277:42669–42679. 2002. View Article : Google Scholar : PubMed/NCBI
32	Yang JW, Kim MR, Kim HG, Kim SK, Jeong HG and Kang KW: Differential regulation of ErbB2 expression by cAMP-dependent protein kinase in tamoxifen-resistant breast cancer cells. Arch Pharm Res. 31:350–356. 2008. View Article : Google Scholar : PubMed/NCBI
33	Barrett JM, Puglia MA, Singh G and Tozer RG: Expression of Ets-related transcription factors and matrix metalloproteinase genes in human breast cancer cells. Breast Cancer Res Treat. 72:227–232. 2002. View Article : Google Scholar : PubMed/NCBI
34	Chotteau-Lelièvre A, Révillion F, Lhotellier V, Hornez L, Desbiens X, Cabaret V, de Launoit Y and Peyrat JP: Prognostic value of ERM gene expression in human primary breast cancers. Clin Cancer Res. 10:7297–7303. 2004. View Article : Google Scholar : PubMed/NCBI
35	Goueli BS and Janknecht R: Upregulation of the catalytic telomerase subunit by the transcription factor ER81 and oncogenic HER2/Neu, Ras, or Raf. Mol Cell Biol. 24:25–35. 2004. View Article : Google Scholar : PubMed/NCBI
36	Jiang J, Wei Y, Liu D, Zhou J, Shen J, Chen X, Zhang S, Kong X and Gu J: E1AF promotes breast cancer cell cycle progression via upregulation of Cyclin D3 transcription. Biochem Biophys Res Commun. 358:53–58. 2007. View Article : Google Scholar : PubMed/NCBI
37	Shepherd TG, Kockeritz L, Szrajber MR, Muller WJ and Hassell JA: The pea3 subfamily ETS genes are required for HER2/Neu-mediated mammary oncogenesis. Curr Biol. 11:1739–1748. 2001. View Article : Google Scholar : PubMed/NCBI
38	Wu X and Sukumar S: ETS genes in breast cancer: A step in the right direction. Cancer Biol Ther. 6:83–84. 2007. View Article : Google Scholar : PubMed/NCBI
39	Hong H, Yu H, Yuan J, Guo C, Cao H, Li W and Xiao C: MicroRNA-200b impacts breast cancer cell migration and invasion by regulating Ezrin-Radixin-Moesin. Med Sci Monit. 22:1946–1952. 2016. View Article : Google Scholar : PubMed/NCBI
40	Baert JL, Monté D, Musgrove EA, Albagli O, Sutherland RL and Launoit Y: Expression of the PEA3 group of ETS-related transcription factors in human breast-cancer cells. Int J Cancer. 70:590–597. 1997. View Article : Google Scholar : PubMed/NCBI
41	Müller P, Kietz S, Gustafsson JA and Ström A: The anti-estrogenic effect of all-trans-retinoic acid on the breast cancer cell line MCF-7 is dependent on HES-1 expression. J Biol Chem. 277:28376–28379. 2002. View Article : Google Scholar : PubMed/NCBI
42	Müller P, Crofts JD, Newman BS, Bridgewater LC, Lin CY, Gustafsson JA and Ström A: SOX9 mediates the retinoic acid-induced HES-1 gene expression in human breast cancer cells. Breast Cancer Res Treat. 120:317–326. 2010. View Article : Google Scholar : PubMed/NCBI
43	Park S-W, Do H-J, Ha WT, Han MH, Song H, Uhm SJ, Chung HJ and Kim JH: Differential expression of ETS family transcription factors in NCCIT human embryonic carcinoma cells upon retinoic acid-induced differentiation. Biol Pharm Bull. 37:659–665. 2014. View Article : Google Scholar : PubMed/NCBI
44	Owman C, Blay P, Nilsson C and Lolait SJ: Cloning of human cDNA encoding a novel heptahelix receptor expressed in Burkitt's lymphoma and widely distributed in brain and peripheral tissues. Biochem Biophys Res Commun. 228:285–292. 1996. View Article : Google Scholar : PubMed/NCBI
45	Olde B and Leeb-Lundberg LM: GPR30/GPER1: Searching for a role in estrogen physiology. Trends Endocrinol Metab. 20:409–416. 2009. View Article : Google Scholar : PubMed/NCBI
46	Zhou X, Wang S, Wang Z, Feng X, Liu P, Lv XB, Li F, Yu FX, Sun Y, Yuan H, et al: Estrogen regulates Hippo signaling via GPER in breast cancer. J Clin Invest. 125:2123–2135. 2015. View Article : Google Scholar : PubMed/NCBI
47	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
48	Lappano R, Pisano A and Maggiolini M: GPER function in breast cancer: An overview. Front Endocrinol (Lausanne). 5:662014. View Article : Google Scholar : PubMed/NCBI
49	Weißenborn C, Ignatov T, Poehlmann A, Wege AK, Costa SD, Zenclussen AC and Ignatov A: GPER functions as a tumor suppressor in MCF-7 and SK-BR-3 breast cancer cells. J Cancer Res Clin Oncol. 140:663–671. 2014. View Article : Google Scholar : PubMed/NCBI
50	Martin SG, Lebot MN, Sukkarn B, Ball G, Green AR, Rakha EA, Ellis IO and Storr SJ: Low expression of G protein-coupled oestrogen receptor 1 (GPER) is associated with adverse survival of breast cancer patients. Oncotarget. 9:25946–25956. 2018. View Article : Google Scholar : PubMed/NCBI
51	Manjegowda MC, Gupta PS and Limaye AM: Hyper-methylation of the upstream CpG island shore is a likely mechanism of GPER1 silencing in breast cancer cells. Gene. 614:65–73. 2017. View Article : Google Scholar : PubMed/NCBI
52	Ignatov T, Weißenborn C, Poehlmann A, Lemke A, Semczuk A, Roessner A, Costa SD, Kalinski T and Ignatov A: GPER-1 expression decreases during breast cancer tumorigenesis. Cancer Invest. 31:309–315. 2013. View Article : Google Scholar : PubMed/NCBI
53	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
54	Lv X, He C, Huang C, Hua G, Wang Z, Remmenga SW, Rodabough KJ, Karpf AR, Dong J, Davis JS and Wang C: G-1 inhibits breast cancer cell growth via targeting colchicine-binding site of tubulin to interfere with microtubule assembly. Mol Cancer Ther. 16:1080–1091. 2017. View Article : Google Scholar : PubMed/NCBI
55	Weissenborn C, Ignatov T, Nass N, Kalinski T, Dan Costa S, Zenclussen AC and Ignatov A: GPER promoter methylation controls GPER expression in breast cancer patients. Cancer Invest. 35:100–107. 2017. View Article : Google Scholar : PubMed/NCBI
56	Kohrman AQ and Matus DQ: Divide or conquer: Cell cycle regulation of invasive behavior. Trends Cell Biol. 27:12–25. 2017. View Article : Google Scholar : PubMed/NCBI
57	Kamath L, Meydani A, Foss F and Kuliopulos A: Signaling from protease-activated receptor-1 inhibits migration and invasion of breast cancer cells. Cancer Res. 61:5933–5940. 2001.PubMed/NCBI
58	Ignatov A, Ignatov T, Roessner A, Costa SD and Kalinski T: Role of GPR30 in the mechanisms of tamoxifen resistance in breast cancer MCF-7 cells. Breast Cancer Res Treat. 123:87–96. 2010. View Article : Google Scholar : PubMed/NCBI
59	Broselid S, Cheng B, Sjöström M, Lövgren K, Klug-De Santiago HL, Belting M, Jirström K, Malmström P, Olde B, Bendahl PO, et al: G protein-coupled estrogen receptor is apoptotic and correlates with increased distant disease-free survival of estrogen receptor-positive breast cancer patients. Clin Cancer Res. 19:1681–1692. 2013. View Article : Google Scholar : PubMed/NCBI
60	Vivacqua A, Lappano R, De Marco P, Sisci D, Aquila S, De Amicis F, Fuqua SA, Andò 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
61	Manjegowda MC and Limaye AM: DNA methylation dependent suppression of GPER1 in colorectal cancer. Med Res Arch. 6:2018.
62	Lelièvre E, Lionneton F, Soncin F and Vandenbunder B: The Ets family contains transcriptional activators and repressors involved in angiogenesis. Int J Biochem Cell Biol. 33:391–407. 2001. View Article : Google Scholar : PubMed/NCBI
63	Mavrothalassitis G and Ghysdael J: Proteins of the ETS family with transcriptional repressor activity. Oncogene. 19:6524–6532. 2000. View Article : Google Scholar : PubMed/NCBI
64	Kim GC, Kwon HK, Lee CG, Verma R, Rudra D, Kim T, Kang K, Nam JH, Kim Y and Im SH: Upregulation of Ets1 expression by NFATc2 and NFKB1/RELA promotes breast cancer cell invasiveness. Oncogenesis. 7:912018. View Article : Google Scholar : PubMed/NCBI
65	Wallace JA, Li F, Balakrishnan S, Cantemir-Stone CZ, Pecot T, Martin C, Kladney RD, Sharma SM, Trimboli AJ, Fernandez SA, et al: Ets2 in tumor fibroblasts promotes angiogenesis in breast cancer. PLoS One. 8:e715332013. View Article : Google Scholar : PubMed/NCBI
66	de Launoit Y, Chotteau-Lelievre A, Beaudoin C, Coutte L, Netzer S, Brenner C, Huvent I and Baert JL: The PEA3 group of ETS-related transcription factors: Role in breast cancer metastasis. Adv Exp Med Biol. 480:107–116. 2000. View Article : Google Scholar : PubMed/NCBI
67	De Luca A, Maiello MR, D'Alessio A, Pergameno M and Normanno N: The RAS/RAF/MEK/ERK and the PI3K/AKT signalling pathways: Role in cancer pathogenesis and implications for therapeutic approaches. Expert Opin Ther Targets. 16 (Suppl 2):S17–S27. 2012. View Article : Google Scholar : PubMed/NCBI
68	Jiang P, Enomoto A and Takahashi M: Cell biology of the movement of breast cancer cells: Intracellular signalling and the actin cytoskeleton. Cancer Lett. 284:122–130. 2009. View Article : Google Scholar : PubMed/NCBI
69	Nguyen DH, Hussaini IM and Gonias SL: Binding of urokinase-type plasminogen activator to its receptor in MCF-7 cells activates extracellular signal-regulated kinase 1 and 2 which is required for increased cellular motility. J Biol Chem. 273:8502–8507. 1998. View Article : Google Scholar : PubMed/NCBI
70	Palorini R, Votta G, Pirola Y, De Vitto H, De Palma S, Airoldi C, Vasso M, Ricciardiello F, Lombardi PP, Cirulli C, et al: Protein kinase A activation promotes cancer cell resistance to glucose starvation and anoikis. PLoS Genet. 12:e10059312016. View Article : Google Scholar : PubMed/NCBI
71	Svensson S, Jirström K, Rydén L, Roos G, Emdin S, Ostrowski MC and Landberg G: ERK phosphorylation is linked to VEGFR2 expression and Ets-2 phosphorylation in breast cancer and is associated with tamoxifen treatment resistance and small tumours with good prognosis. Oncogene. 24:4370–4379. 2005. View Article : Google Scholar : PubMed/NCBI
72	Switzer CH, Cheng RY, Ridnour LA, Glynn SA, Ambs S and Wink DA: Ets-1 is a transcriptional mediator of oncogenic nitric oxide signaling in estrogen receptor-negative breast cancer. Breast Cancer Res. 14:R1252012. View Article : Google Scholar : PubMed/NCBI
73	Baert J-L, Beaudoin C, Coutte L and De Launoit Y: ERM transactivation is up-regulated by the repression of DNA binding after the PKA phosphorylation of a consensus site at the edge of the ETS domain. J Biol Chem. 277:1002–1012. 2002. View Article : Google Scholar : PubMed/NCBI
74	Keld R, Guo B, Downey P, Cummins R, Gulmann C, Ang YS and Sharrocks AD: PEA3/ETV4-related transcription factors coupled with active ERK signalling are associated with poor prognosis in gastric adenocarcinoma. Br J Cancer. 105:124–130. 2011. View Article : Google Scholar : PubMed/NCBI