G-protein-coupled estrogen receptor suppresses the migration of osteosarcoma cells via post-translational regulation of Snail

  • Zhen Wang
  • Xiao Chen
  • Yongqiang Zhao
  • Yi Jin
  • Jia ZhengEmail author
Original Article – Cancer Research



Emerging evidences show that G-protein-coupled estrogen receptor (GPER) can regulate the progression of various cancers, while its roles in the progression of osteosarcoma (OS) are not well illustrated.


The expression of GPER in OS cells and tissues were checked. Its roles in cell migration and expression of Snail was checked by use of its agonist G-1.


We found that the expression of GPER in OS cells and tissues were lower than that in their corresponding controls. OS patients with higher levels of GPER showed increased overall survival rate (OS) as compared with the lower ones. The activator of GPER (G-1) or overexpression of GPER can inhibit the migration and invasion of OS cells and downregulate mesenchymal markers. G-1 can rapidly decrease the expression of Snail, one powerful epithelial–mesenchymal transition transcription factor (EMT-TF). Overexpression of Snail can attenuate the suppression effects of G-1 on migration of OS cells, suggesting that Snail was involved in GPER-regulated migration of OS cells. Mechanically, G-1 rapidly decreased the protein of Snail but had no effect on its mRNA expression. This was because G-1 can decrease the protein stability of Snail. Further, G-1 increased the expression of FBXL5, which can trigger the proteasome-mediated degradation of Snail. Knockdown of FBXL5 can reverse G-1-induced downregulation of Snail in OS cells.


Activation of GPER can suppress the migration and invasion of OS cells via FBXL5-mediated post-translational down regulation of Snail. It suggested that targeted activation of GPER might be a potent potential therapy approach to overcome the metastasis of OS patients.


GPER Snail EMT Osteosarcoma FBXL5 


Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Research involving human participants and/or animals

No human or animal study.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. Anderson ME (2016) Update on survival in osteosarcoma. Orthop Clin N Am 47:283–292CrossRefGoogle Scholar
  2. Azmi AS, Muqbil I, Wu J, Aboukameel A, Senapedis W, Baloglu E, Bollig-Fischer A, Dyson G, Kauffman M, Landesman Y et al (2015) Targeting the nuclear export protein XPO1/CRM1 reverses epithelial to mesenchymal transition. Sci Rep 5:16077CrossRefGoogle Scholar
  3. Brabletz T, Kalluri R, Nieto MA, Weinberg RA (2018) EMT in cancer. Nat Rev Cancer 18:128–128+CrossRefGoogle Scholar
  4. Bustos V, Nolan AM, Nijhuis A, Harvey H, Parker A, Poulsom R, McBryan J, Thomas W, Silver A, Harvey BJ (2017) GPER mediates differential effects of estrogen on colon cancer cell proliferation and migration under normoxic and hypoxic conditions. Oncotarget 8:84258–84275CrossRefGoogle Scholar
  5. Chen P, Wang H, Duan Z, Zou JX, Chen H, He W, Wang J (2014) Estrogen-related receptor alpha confers methotrexate resistance via attenuation of reactive oxygen species production and P53 mediated apoptosis in osteosarcoma cells. Biomed Res Int 2014:616025Google Scholar
  6. Chen Y, Zhang K, Li Y, He Q (2017) Estrogen-related receptor alpha participates transforming growth factor-beta (TGF-beta) induced epithelial-mesenchymal transition of osteosarcoma cells. Cell Adhes Migr 11:338–346CrossRefGoogle Scholar
  7. Chou AJ, Geller DS, Gorlick R (2008) Therapy for osteosarcoma: where do we go from here? Paediatr Drugs 10:315–327CrossRefGoogle Scholar
  8. Daw NC, Chou AJ, Jaffe N, Rao BN, Billups CA, Rodriguez-Galindo C, Meyers PA, Huh WW (2015) Recurrent osteosarcoma with a single pulmonary metastasis: a multi-institutional review. Br J Cancer 112:278–282CrossRefGoogle Scholar
  9. Fang D, Yang H, Lin J, Teng Y, Jiang Y, Chen J, Li Y (2015) 17beta-estradiol regulates cell proliferation, colony formation, migration, invasion and promotes apoptosis by upregulating miR-9 and thus degrades MALAT-1 in osteosarcoma cell MG-63 in an estrogen receptor-independent manner. Biochem Biophys Res Commun 457:500–506CrossRefGoogle Scholar
  10. Fang S, Yu L, Mei H, Yang J, Gao T, Cheng A, Guo W, Xia K, Liu G (2016) Cisplatin promotes mesenchymal-like characteristics in osteosarcoma through Snail. Oncol Lett 12:5007–5014CrossRefGoogle Scholar
  11. Feldman RD, Ding Q, Hussain Y, Limbird LE, Pickering JG, Gros R (2016) Aldosterone mediates metastatic spread of renal cancer via the G protein-coupled estrogen receptor (GPER). FASEB J 30:2086–2096CrossRefGoogle Scholar
  12. Fic A, Mlakar SJ, Juvan P, Mlakar V, Marc J, Dolenc MS, Broberg K, Masic LP (2015) Genome-wide gene expression profiling of low-dose, long-term exposure of human osteosarcoma cells to bisphenol A and its analogs bisphenols AF and S. Toxicol In Vitro 29:1060–1069CrossRefGoogle Scholar
  13. Filardo EJ (2018) A role for G-protein coupled estrogen receptor (GPER) in estrogen-induced carcinogenesis: dysregulated glandular homeostasis, survival and metastasis. J Steroid Biochem Mol Biol 176:38–48CrossRefGoogle Scholar
  14. Filardo EJ, Quinn JA, Bland KI, Frackelton AR Jr (2000) Estrogen-induced activation of Erk-1 and Erk-2 requires the G protein-coupled receptor homolog, GPR30, and occurs via trans-activation of the epidermal growth factor receptor through release of HB-EGF. Mol Endocrinol (Baltim MD) 14:1649–1660CrossRefGoogle Scholar
  15. Hattinger CM, Fanelli M, Tavanti E, Vella S, Ferrari S, Picci P, Serra M (2015) Advances in emerging drugs for osteosarcoma. Expert Opin Emerg Drugs 20:495–514CrossRefGoogle Scholar
  16. Kansara M, Teng MW, Smyth MJ, Thomas DM (2014) Translational biology of osteosarcoma. Nat Rev Cancer 14:722–735CrossRefGoogle Scholar
  17. Liang S, Chen Z, Jiang G, Zhou Y, Liu Q, Su Q, Wei W, Du J, Wang H (2017) Activation of GPER suppresses migration and angiogenesis of triple negative breast cancer via inhibition of NF-kappaB/IL-6 signals. Cancer Lett 386:12–23CrossRefGoogle Scholar
  18. Maggiolini M, Picard D (2010) The unfolding stories of GPR30, a new membrane-bound estrogen receptor. J Endocrinol 204:105–114CrossRefGoogle Scholar
  19. Marjon NA, Hu C, Hathaway HJ, Prossnitz ER (2014) G protein-coupled estrogen receptor regulates mammary tumorigenesis and metastasis. Mol Cancer Res 12:1644–1654CrossRefGoogle Scholar
  20. Meazza C, Scanagatta P (2016) Metastatic osteosarcoma: a challenging multidisciplinary treatment. Expert Rev Anticancer 16:543–556CrossRefGoogle Scholar
  21. Meng X, Zhu Y, Tao L, Zhao S, Qiu S (2018) miR-590-3p mediates melatonin-induced cell apoptosis by targeting septin 7 in the human osteoblast cell line hFOB 1.19. Mol Med Rep 17:7202–7208Google Scholar
  22. Molina L, Figueroa CD, Bhoola KD, Ehrenfeld P (2017) GPER-1/GPR30 a novel estrogen receptor sited in the cell membrane: therapeutic coupling to breast cancer. Expert Opin Ther Targets 21:755–766CrossRefGoogle Scholar
  23. Nieto MA (2002) The snail superfamily of zinc-finger transcription factors. Nat Rev Mol Cell Biol 3:155–166CrossRefGoogle Scholar
  24. Revankar CM, Cimino DF, Sklar LA, Arterburn JB, Prossnitz ER (2005) A transmembrane intracellular estrogen receptor mediates rapid cell signaling. Science 307:1625–1630CrossRefGoogle Scholar
  25. Salvatori L, Caporuscio F, Coroniti G, Starace G, Frati L, Russo MA, Petrangeli E (2009) Down-regulation of epidermal growth factor receptor induced by estrogens and phytoestrogens promotes the differentiation of U2OS human osteosarcoma cells. J Cell Physiol 220:35–44CrossRefGoogle Scholar
  26. Svoboda M, Hamilton G, Thalhammer T (2010) Steroid hormone metabolizing enzymes in benign and malignant human bone tumors. Expert Opin Drug Metab Toxicol 6:427–437CrossRefGoogle Scholar
  27. Tang Z, Li C, Kang B, Gao G, Li C, Zhang Z (2017) GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses. Nucleic Acids Res 45:W98–W102CrossRefGoogle Scholar
  28. Vinas-Castells R, Beltran M, Valls G, Gomez I, Garcia JM, Montserrat-Sentis B, Baulida J, Bonilla F, de Herreros AG, Diaz VM (2010) The hypoxia-controlled FBXL14 ubiquitin ligase targets SNAIL1 for proteasome degradation. J Biol Chem 285:3794–3805CrossRefGoogle Scholar
  29. Vinas-Castells R, Frias A, Robles-Lanuza E, Zhang K, Longmore GD, Garcia de Herreros A, Diaz VM (2014) Nuclear ubiquitination by FBXL5 modulates Snail1 DNA binding and stability. Nucleic Acids Res 42:1079–1094CrossRefGoogle Scholar
  30. Wei W, Chen ZJ, Zhang KS, Yang XL, Wu YM, Chen XH, Huang HB, Liu HL, Cai SH, Du J et al (2014) 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:e1428CrossRefGoogle Scholar
  31. Yang M, Liu B, Jin L, Tao H, Yang Z (2017) Estrogen receptor beta exhibited anti-tumor effects on osteosarcoma cells by regulating integrin, IAP, NF-kB/BCL-2 and PI3K/Akt signal pathway. J Bone Oncol 9:15–20CrossRefGoogle Scholar
  32. Yen M-L, Chien C-C, Chiu I-M, Huang H-I, Chen Y-C, Hu H-I, Yen BL (2007) Multilineage differentiation and characterization of the human fetal osteoblastic 1.19 cell line: a possible in vitro model of human mesenchymal progenitors. Stem Cells (Dayt Ohio) 25:125–131CrossRefGoogle Scholar
  33. Zheng H, Shen M, Zha YL, Li W, Wei Y, Blanco MA, Ren G, Zhou T, Storz P, Wang HY et al (2014) PKD1 phosphorylation-dependent degradation of SNAIL by SCF-FBXO11 regulates epithelial-mesenchymal transition and metastasis. Cancer Cell 26:358–373CrossRefGoogle Scholar
  34. Zhou BP, Deng J, Xia W, Xu J, Li YM, Gunduz M, Hung MC (2004) Dual regulation of Snail by GSK-3beta-mediated phosphorylation in control of epithelial-mesenchymal transition. Nat Cell Biol 6:931–940CrossRefGoogle Scholar
  35. Zhu G, Huang Y, Wu C, Wei D, Shi Y (2016) Activation of G-protein-coupled estrogen receptor inhibits the migration of human nonsmall cell lung cancer cells via IKK-beta/NF-kappaB signals. DNA Cell Biol 35:434–442CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of OrthopaedicsHenan Provincial People’s HospitalZhengzhouChina

Personalised recommendations