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Breast Cancer Research and Treatment

, Volume 174, Issue 1, pp 65–78 | Cite as

GGNBP2 suppresses triple-negative breast cancer aggressiveness through inhibition of IL-6/STAT3 signaling activation

  • Jingjing Liu
  • Lei Liu
  • Ernesto Yagüe
  • Qianxi Yang
  • Teng Pan
  • Hui Zhao
  • Yunhui HuEmail author
  • Jin ZhangEmail author
Preclinical study
  • 114 Downloads

Abstract

Background

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, lacking effective targeted therapies, and whose underlying mechanisms are still unclear. The gene coding for Gametogenetin-binding protein (GGNBP2), also known as Zinc Finger Protein 403 (ZNF403), is located on chromosome 17q12-q23, a region known as a breast cancer susceptibility locus. We have previously reported that GGNBP2 functions as a tumor suppressor in estrogen receptor-positive breast cancer. The aim of this study was to evaluate the role and mechanisms of GGNBP2 in TNBC.

Methods

The effect of GGNBP2 on TNBC aggressiveness was investigated both in vitro and in vivo. The protein and mRNA expression levels were analyzed by western blotting and reverse transcription quantitative polymerase chain reaction, respectively. Fluorescence-activated cell sorting analysis was used to evaluate the cell cycle distribution and cell apoptosis. Immunohistochemistry was used to determine the expression of GGNBP2 in breast cancer tissues.

Results

We find that GGNBP2 expression decreases in TNBC tissues and is associated with the outcome of breast cancer patients. Furthermore, experimental overexpression of GGNBP2 in MDA-MB-231 and Cal51 cells suppresses cell proliferation, migration and invasion, reduces the cancer stem cell subpopulation, and promotes cell apoptosis in vitro as well as inhibits tumor growth in vivo. In these cell models, overexpression of GGNBP2 decreases the activation of IL-6/STAT3 signaling.

Conclusion

Our data demonstrate that GGNBP2 suppresses cancer aggressiveness by inhibition of IL-6/STAT3 activation in TNBC.

Keywords

Triple-negative breast cancer STAT3 GGNBP2 

Notes

Acknowledgements

This study was supported by the Tianjin Natural Sciences Foundation (17JCQNJC09900 to YH) and the National Natural Science Foundation of China (No. 81672623 to ZJ). EY thanks Breast Cancer Now for supporting research in his laboratory.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

10549_2018_5052_MOESM1_ESM.docx (1.7 mb)
Supplementary material 1 (DOCX 1.8 mb)

References

  1. 1.
    Siegel RL, Miller KD, Jemal A (2018) Cancer statistics, 2018. CA Cancer J Clin 68(1):7–30.  https://doi.org/10.3322/caac.21442 Google Scholar
  2. 2.
    Wein L, Luen SJ, Savas P, Salgado R, Loi S (2018) Checkpoint blockade in the treatment of breast cancer: current status and future directions. Br J Cancer.  https://doi.org/10.1038/s41416-018-0126-6 Google Scholar
  3. 3.
    Ohbayashi T, Oikawa K, Iwata R, Kameta A, Evine K, Isobe T, Matsuda Y, Mimura J, Fujii-Kuriyama Y, Kuroda M, Mukai K (2001) Dioxin induces a novel nuclear factor, DIF-3, that is implicated in spermatogenesis. FEBS Lett 508(3):341–344Google Scholar
  4. 4.
    Glynn RW, Miller N, Kerin MJ (2010) 17q12-21 - the pursuit of targeted therapy in breast cancer. Cancer Treat Rev 36(3):224–229.  https://doi.org/10.1016/j.ctrv.2009.12.007 Google Scholar
  5. 5.
    Cino EA, Choy WY, Karttunen M (2016) Characterization of the free state ensemble of the CoRNR box motif by molecular dynamics simulations. J Phys Chem B 120(6):1060–1068.  https://doi.org/10.1021/acs.jpcb.5b11565 Google Scholar
  6. 6.
    Hu X, Lazar MA (1999) The CoRNR motif controls the recruitment of corepressors by nuclear hormone receptors. Nature 402(6757):93–96.  https://doi.org/10.1038/47069 Google Scholar
  7. 7.
    Zhang J, Wang Y, Zhou Y, Cao Z, Huang P, Lu B (2005) Yeast two-hybrid screens imply that GGNBP1, GGNBP2 and OAZ3 are potential interaction partners of testicular germ cell-specific protein GGN1. FEBS Lett 579(2):559–566.  https://doi.org/10.1016/j.febslet.2004.10.112 Google Scholar
  8. 8.
    Chen A, Li J, Song L, Ji C, Boing M, Chen J, Brand-Saberi B (2017) GGNBP2 is necessary for testis morphology and sperm development. Sci Rep 7(1):2998.  https://doi.org/10.1038/s41598-017-03193-y Google Scholar
  9. 9.
    Li S, Moore AK, Zhu J, Li X, Zhou H, Lin J, He Y, Xing F, Pan Y, Bohler HC, Ding J, Cooney AJ, Lan Z, Lei Z (2016) Ggnbp2 is essential for pregnancy success via regulation of mouse trophoblast stem cell proliferation and differentiation. Biol Reprod 94(2):41.  https://doi.org/10.1095/biolreprod.115.136358 Google Scholar
  10. 10.
    Li Y, Chen Z (2004) Molecular cloning and characterization of LCRG1 a novel gene localized to the tumor suppressor locus D17S800–D17S930. Cancer Lett 209(1):75–85.  https://doi.org/10.1016/j.canlet.2003.11.034 Google Scholar
  11. 11.
    Zhu Z, Lou C, Zheng Z, Zhu R, Tian S, Xie C, Zhao H (2017) ZFP403, a novel tumor suppressor, inhibits the proliferation and metastasis in ovarian cancer. Gynecol Oncol 147(2):418–425.  https://doi.org/10.1016/j.ygyno.2017.08.025 Google Scholar
  12. 12.
    Yin F, Liu L, Liu X, Li G, Zheng L, Li D, Wang Q, Zhang W, Li L (2014) Downregulation of tumor suppressor gene ribonuclease T2 and gametogenetin binding protein 2 is associated with drug resistance in ovarian cancer. Oncol Rep 32(1):362–372.  https://doi.org/10.3892/or.2014.3175 Google Scholar
  13. 13.
    Zhan A, Lei B, Wu H, Wen Y, Zheng L, Wang S, Wan X, Wei Z (2017) GGNBP2 suppresses the proliferation, invasion, and migration of human glioma cells. Oncol Res 25(5):831–842.  https://doi.org/10.3727/096504016X14816726393937 Google Scholar
  14. 14.
    Guan R, Wen XY, Wu J, Duan R, Cao H, Lam S, Hou D, Wang Y, Hu J, Chen Z (2012) Knockdown of ZNF403 inhibits cell proliferation and induces G2/M arrest by modulating cell-cycle mediators. Mol Cell Biochem 365(1–2):211–222.  https://doi.org/10.1007/s11010-012-1262-6 Google Scholar
  15. 15.
    Lan ZJ, Hu Y, Zhang S, Li X, Zhou H, Ding J, Klinge CM, Radde BN, Cooney AJ, Zhang J, Lei Z (2016) GGNBP2 acts as a tumor suppressor by inhibiting estrogen receptor alpha activity in breast cancer cells. Breast Cancer Res Treat 158(2):263–276.  https://doi.org/10.1007/s10549-016-3880-2 Google Scholar
  16. 16.
    Hu Y, Yague E, Zhao J, Wang L, Bai J, Yang Q, Pan T, Zhao H, Liu J, Zhang J (2018) Sabutoclax, pan-active BCL-2 protein family antagonist, overcomes drug resistance and eliminates cancer stem cells in breast cancer. Cancer Lett 423:47–59.  https://doi.org/10.1016/j.canlet.2018.02.036 Google Scholar
  17. 17.
    Azim HA Jr, Peccatori FA, Brohee S, Branstetter D, Loi S, Viale G, Piccart M, Dougall WC, Pruneri G, Sotiriou C (2015) RANK-ligand (RANKL) expression in young breast cancer patients and during pregnancy. Breast Cancer Res BCR 17:24.  https://doi.org/10.1186/s13058-015-0538-7 Google Scholar
  18. 18.
    Hu Y, Li K, Asaduzzaman M, Cuella R, Shi H, Raguz S, Coombes RC, Zhou Y, Yague E (2016) MiR-106b ~ 25 cluster regulates multidrug resistance in an ABC transporter-independent manner via downregulation of EP300. Oncol Rep 35(2):1170–1178.  https://doi.org/10.3892/or.2015.4412 Google Scholar
  19. 19.
    Hu Y, Qiu Y, Yague E, Ji W, Liu J, Zhang J (2016) miRNA-205 targets VEGFA and FGF2 and regulates resistance to chemotherapeutics in breast cancer. Cell Death Dis 7(6):e2291.  https://doi.org/10.1038/cddis.2016.194 Google Scholar
  20. 20.
    Clevers H (2011) The cancer stem cell: premises, promises and challenges. Nat Med 17(3):313–319.  https://doi.org/10.1038/nm.2304 Google Scholar
  21. 21.
    Wang Z, Kong J, Wu Y, Zhang J, Wang T, Li N, Fan J, Wang H, Zhang J, Ling R (2018) PRMT5 determines the sensitivity to chemotherapeutics by governing stemness in breast cancer. Breast Cancer Res Treat 168(2):531–542.  https://doi.org/10.1007/s10549-017-4597-6 Google Scholar
  22. 22.
    Zhou Z, Li M, Zhang L, Zhao H, Sahin O, Chen J, Zhao JJ, Songyang Z, Yu D (2018) Oncogenic kinase-induced PKM2 tyrosine 105 phosphorylation converts nononcogenic PKM2 to a tumor promoter and induces cancer stem-like cells. Cancer Res 78(9):2248–2261.  https://doi.org/10.1158/0008-5472.CAN-17-2726 Google Scholar
  23. 23.
    Suman S, Das TP, Damodaran C (2013) Silencing NOTCH signaling causes growth arrest in both breast cancer stem cells and breast cancer cells. Br J Cancer 109(10):2587–2596.  https://doi.org/10.1038/bjc.2013.642 Google Scholar
  24. 24.
    Asaduzzaman M, Constantinou S, Min H, Gallon J, Lin ML, Singh P, Raguz S, Ali S, Shousha S, Coombes RC, Lam EW, Hu Y, Yague E (2017) Tumour suppressor EP300, a modulator of paclitaxel resistance and stemness, is downregulated in metaplastic breast cancer. Breast Cancer Res Treat 163(3):461–474.  https://doi.org/10.1007/s10549-017-4202-z Google Scholar
  25. 25.
    Hu Y, Guo R, Wei J, Zhou Y, Ji W, Liu J, Zhi X, Zhang J (2015) Effects of PI3K inhibitor NVP-BKM120 on overcoming drug resistance and eliminating cancer stem cells in human breast cancer cells. Cell Death Dis 6:e2020.  https://doi.org/10.1038/cddis.2015.363 Google Scholar
  26. 26.
    Balko JM, Schwarz LJ, Bhola NE, Kurupi R, Owens P, Miller TW, Gomez H, Cook RS, Arteaga CL (2013) Activation of MAPK pathways due to DUSP4 loss promotes cancer stem cell-like phenotypes in basal-like breast cancer. Cancer Res 73(20):6346–6358.  https://doi.org/10.1158/0008-5472.CAN-13-1385 Google Scholar
  27. 27.
    Kim SY, Kang JW, Song X, Kim BK, Yoo YD, Kwon YT, Lee YJ (2013) Role of the IL-6-JAK1-STAT3-Oct-4 pathway in the conversion of non-stem cancer cells into cancer stem-like cells. Cell Signal 25(4):961–969.  https://doi.org/10.1016/j.cellsig.2013.01.007 Google Scholar
  28. 28.
    Yanai A, Inoue N, Yagi T, Nishimukai A, Miyagawa Y, Murase K, Imamura M, Enomoto Y, Takatsuka Y, Watanabe T, Hirota S, Sasa M, Katagiri T, Miyoshi Y (2015) Activation of mTOR/S6K but not MAPK pathways might be associated with high Ki-67, ER(+), and HER2(–) breast cancer. Clin Breast Cancer 15(3):197–203.  https://doi.org/10.1016/j.clbc.2014.12.002 Google Scholar
  29. 29.
    Marotta LL, Almendro V, Marusyk A, Shipitsin M, Schemme J, Walker SR, Bloushtain-Qimron N, Kim JJ, Choudhury SA, Maruyama R, Wu Z, Gonen M, Mulvey LA, Bessarabova MO, Huh SJ, Silver SJ, Kim SY, Park SY, Lee HE, Anderson KS, Richardson AL, Nikolskaya T, Nikolsky Y, Liu XS, Root DE, Hahn WC, Frank DA, Polyak K (2011) The JAK2/STAT3 signaling pathway is required for growth of CD44(+)CD24(–) stem cell-like breast cancer cells in human tumors. J Clin Invest 121(7):2723–2735.  https://doi.org/10.1172/JCI44745 Google Scholar
  30. 30.
    Couture LA, Harris MW, Birnbaum LS (1990) Characterization of the peak period of sensitivity for the induction of hydronephrosis in C57BL/6N mice following exposure to 2,3,7, 8-tetrachlorodibenzo-p-dioxin. Fundam Appl Toxicol 15(1):142–150Google Scholar
  31. 31.
    Reuter TY, Medhurst AL, Waisfisz Q, Zhi Y, Herterich S, Hoehn H, Gross HJ, Joenje H, Hoatlin ME, Mathew CG, Huber PA (2003) Yeast two-hybrid screens imply involvement of Fanconi anemia proteins in transcription regulation, cell signaling, oxidative metabolism, and cellular transport. Exp Cell Res 289(2):211–221Google Scholar
  32. 32.
    Lv W, Su B, Li Y, Geng C, Chen N (2018) KIAA0101 inhibition suppresses cell proliferation and cell cycle progression by promoting the interaction between p53 and Sp1 in breast cancer. Biochem Biophys Res Commun.  https://doi.org/10.1016/j.bbrc.2018.06.046 Google Scholar
  33. 33.
    Wang X, Li L, Wu Y, Zhang R, Zhang M, Liao D, Wang G, Qin G, Xu RH, Kang T (2016) CBX4 suppresses metastasis via recruitment of HDAC3 to the Runx2 promoter in colorectal carcinoma. Cancer Res 76(24):7277–7289.  https://doi.org/10.1158/0008-5472.CAN-16-2100 Google Scholar
  34. 34.
    Zhou Y, Hu Y, Yang M, Jat P, Li K, Lombardo Y, Xiong D, Coombes RC, Raguz S, Yague E (2014) The miR-106b ~ 25 cluster promotes bypass of doxorubicin-induced senescence and increase in motility and invasion by targeting the E-cadherin transcriptional activator EP300. Cell Death Differ 21(3):462–474.  https://doi.org/10.1038/cdd.2013.167 Google Scholar
  35. 35.
    Garcia-Carpizo V, Ruiz-Llorente S, Sarmentero J, Grana-Castro O, Pisano DG, Barrero MJ (2018) CREBBP/EP300 bromodomains are critical to sustain the GATA1/MYC regulatory axis in proliferation. Epigenet Chromatin 11(1):30.  https://doi.org/10.1186/s13072-018-0197-x Google Scholar
  36. 36.
    Attar N, Kurdistani SK (2017) Exploitation of EP300 and CREBBP lysine acetyltransferases by cancer. Cold Spring Harb Perspect Med 7 (3).  https://doi.org/10.1101/cshperspect.a026534
  37. 37.
    Kong X, Zhang J, Li J, Shao J, Fang L (2018) MiR-130a-3p inhibits migration and invasion by regulating RAB5B in human breast cancer stem cell-like cells. Biochem Biophys Res Commun 501(2):486–493.  https://doi.org/10.1016/j.bbrc.2018.05.018 Google Scholar
  38. 38.
    Liu M, Liu Y, Deng L, Wang D, He X, Zhou L, Wicha MS, Bai F, Liu S (2018) Transcriptional profiles of different states of cancer stem cells in triple-negative breast cancer. Mol Cancer 17(1):65.  https://doi.org/10.1186/s12943-018-0809-x Google Scholar
  39. 39.
    Yu Y, Luo W, Yang ZJ, Chi JR, Li YR, Ding Y, Ge J, Wang X, Cao XC (2018) miR-190 suppresses breast cancer metastasis by regulation of TGF-beta-induced epithelial-mesenchymal transition. Mol Cancer 17(1):70.  https://doi.org/10.1186/s12943-018-0818-9 Google Scholar
  40. 40.
    Dethlefsen C, Hojfeldt G, Hojman P (2013) The role of intratumoral and systemic IL-6 in breast cancer. Breast Cancer Res Treat 138(3):657–664.  https://doi.org/10.1007/s10549-013-2488-z Google Scholar
  41. 41.
    Schafer ZT, Brugge JS (2007) IL-6 involvement in epithelial cancers. J Clin Invest 117(12):3660–3663.  https://doi.org/10.1172/JCI34237 Google Scholar
  42. 42.
    Guo Y, Xu F, Lu T, Duan Z, Zhang Z (2012) Interleukin-6 signaling pathway in targeted therapy for cancer. Cancer Treat Rev 38(7):904–910.  https://doi.org/10.1016/j.ctrv.2012.04.007 Google Scholar
  43. 43.
    Sherry MM, Reeves A, Wu JK, Cochran BH (2009) STAT3 is required for proliferation and maintenance of multipotency in glioblastoma stem cells. Stem Cells 27(10):2383–2392.  https://doi.org/10.1002/stem.185 Google Scholar
  44. 44.
    Choi HS, Kim JH, Kim SL, Deng HY, Lee D, Kim CS, Yun BS, Lee DS (2018) Catechol derived from aronia juice through lactic acid bacteria fermentation inhibits breast cancer stem cell formation via modulation Stat3/IL-6 signaling pathway. Mol Carcinog.  https://doi.org/10.1002/mc.22870 Google Scholar
  45. 45.
    Gyamfi J, Lee YH, Eom M, Choi J (2018) Interleukin-6/STAT3 signalling regulates adipocyte induced epithelial-mesenchymal transition in breast cancer cells. Sci Rep 8(1):8859.  https://doi.org/10.1038/s41598-018-27184-9 Google Scholar
  46. 46.
    Beebe JD, Liu JY, Zhang JT (2018) Two decades of research in discovery of anticancer drugs targeting STAT3, how close are we? Pharmacol Ther.  https://doi.org/10.1016/j.pharmthera.2018.06.006 Google Scholar
  47. 47.
    Zhang J, Liang Q, Lei Y, Yao M, Li L, Gao X, Feng J, Zhang Y, Gao H, Liu DX, Lu J, Huang B (2012) SOX4 induces epithelial-mesenchymal transition and contributes to breast cancer progression. Cancer Res 72(17):4597–4608.  https://doi.org/10.1158/0008-5472.CAN-12-1045 Google Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.The 3rd Department of Breast Cancer, Treatment and Research Center, China Tianjin Breast Cancer Prevention, Tianjin Medical University Cancer Institute and HospitalNational Clinical Research Center of CancerTianjinPeople’s Republic of China
  2. 2.Division of Cancer, Faculty of Medicine, Cancer Research CenterImperial College LondonLondonUK

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