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Androgen dihydrotestosterone (DHT) promotes the bladder cancer nuclear AR-negative cell invasion via a newly identified membrane androgen receptor (mAR-SLC39A9)-mediated Gαi protein/MAPK/MMP9 intracellular signaling

Oncogene volume 39, pages 574–586 (2020)Cite this article

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Abstract

While androgens may function via nuclear androgen receptor (nAR) to increase bladder cancer (BCa) progression, the impact of androgens on muscle invasive BCa, which contains nearly 80% nAR-negative cells, remains unclear. To dissect the androgens potential impacts on these nAR-negative muscle invasive BCa, we first found that the androgens, dihydrotestosterone (DHT) might function via a novel membrane AR (mAR-SLC39A9) to increase nAR-negative BCa cell migration and invasion. Mechanism dissection revealed that DHT/mAR-SLC39A9 might function by altering Gαi protein-mediated MAPK/MMP9 intracellular signaling to increase nAR-negative BCa cell migration and invasion. Preclinical studies using multiple in vitro nAR-negative BCa cell lines and an in vivo mouse model all demonstrated that targeting this newly identified DHT/mAR-SLC39A9/Gαi/MAPK/MMP9 signaling with small molecules mAR-SLC39A9-shRNA or Gαi-shRNA, and not the classic antiandrogens including enzalutamide, bicalutamide, or hydroxyflutamide, could suppress nAR-negative BCa cell invasion. Results from human clinical samples surveys also indicated the positive correlation of this newly identified DHT/mAR signaling with BCa progression and prognosis. Together, these results suggest that androgens may not only function via the classic nAR to increase the nAR-positive BCa cell invasion, they may also function via this newly identified mAR-SLC39A9 to increase the nAR-negative/mAR-positive BCa cell invasion.

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References

  1. Antoni S, Ferlay J, Soerjomataram I, Znaor A, Jemal A, Bray F. Bladder cancer incidence and mortality: a global overview and recent trends. Eur Urol. 2017;71:96–108.

    Article  PubMed  Google Scholar 

  2. Siegel RL, Miller KD, Jemal A. Cancer Statistics, 2017. CA Cancer J Clin. 2017;67:7–30.

    PubMed  Google Scholar 

  3. Kamat AM, Hahn NM, Efstathiou JA, Lerner SP, Malmstrom PU, Choi W, et al. Bladder cancer. Lancet. 2016;388:2796–810.

    Article  PubMed  Google Scholar 

  4. Tan WS, Rodney S, Lamb B, Feneley M, Kelly J. Management of non-muscle invasive bladder cancer: a comprehensive analysis of guidelines from the United States, Europe and Asia. Cancer Treat Rev. 2016;47:22–31.

    Article  PubMed  Google Scholar 

  5. Milowsky MI, Rumble RB, Booth CM, Gilligan T, Eapen LJ, Hauke RJ, et al. Guideline on muscle-invasive and metastatic bladder cancer (European Association of Urology Guideline): American Society of Clinical Oncology Clinical Practice Guideline Endorsement. J Clin Oncol. 2016;34:1945–52.

    Article  PubMed  Google Scholar 

  6. Carneiro BA, Meeks JJ, Kuzel TM, Scaranti M, Abdulkadir SA, Giles FJ. Emerging therapeutic targets in bladder cancer. Cancer Treat Rev. 2015;41:170–8.

    Article  CAS  PubMed  Google Scholar 

  7. Dobruch J, Daneshmand S, Fisch M, Lotan Y, Noon AP, Resnick MJ, et al. Gender and bladder cancer: a collaborative review of etiology, biology, and outcomes. Eur Urol. 2016;69:300–10.

    Article  PubMed  Google Scholar 

  8. Siegel R, Ward E, Brawley O, Jemal A. Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin. 2011;61:212–36.

    Article  PubMed  Google Scholar 

  9. Chang C, Lee SO, Yeh S, Chang TM. Androgen receptor (AR) differential roles in hormone-related tumors including prostate, bladder, kidney, lung, breast and liver. Oncogene. 2014;33:3225–34.

    Article  CAS  PubMed  Google Scholar 

  10. Chang CS, Kokontis J, Liao ST. Molecular cloning of human and rat complementary DNA encoding androgen receptors. Science. 1988;240:324–6.

    Article  CAS  PubMed  Google Scholar 

  11. Li P, Chen J, Miyamoto H. Androgen receptor signaling in bladder cancer. Cancers (Basel). 2017;9:E20.

    Article  Google Scholar 

  12. Miyamoto H, Zheng Y, Izumi K. Nuclear hormone receptor signals as new therapeutic targets for urothelial carcinoma. Curr Cancer Drug Targets. 2012;12:14–22.

    Article  CAS  PubMed  Google Scholar 

  13. Ou Z, Wang Y, Liu L, Li L, Yeh S, Qi L, et al. Tumor microenvironment B cells increase bladder cancer metastasis via modulation of the IL-8/androgen receptor (AR)/MMPs signals. Oncotarget. 2015;6:26065–78.

    PubMed  PubMed Central  Google Scholar 

  14. Tuygun C, Kankaya D, Imamoglu A, Sertcelik A, Zengin K, Oktay M, et al. Sex-specific hormone receptors in urothelial carcinomas of the human urinary bladder: a comparative analysis of clinicopathological features and survival outcomes according to receptor expression. Urol Oncol. 2011;29:43–51.

    Article  CAS  PubMed  Google Scholar 

  15. Ide H, Inoue S, Miyamoto H. Histopathological and prognostic significance of the expression of sex hormone receptors in bladder cancer: a meta-analysis of immunohistochemical studies. PLoS ONE. 2017;12:e0174746.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Sun YH, Gao X, Tang YJ, Xu CL, Wang LH. Androgens induce increases in intracellular calcium via a G protein-coupled receptor in LNCaP prostate cancer cells. J Androl. 2006;27:671–8.

    Article  CAS  PubMed  Google Scholar 

  17. Sen A, Prizant H, Hammes SR. Understanding extranuclear (nongenomic) androgen signaling: what a frog oocyte can tell us about human biology. Steroids. 2011;76:822–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Foradori CD, Weiser MJ, Handa RJ. Non-genomic actions of androgens. Front Neuroendocr. 2008;29:169–81.

    Article  CAS  Google Scholar 

  19. Miyamoto H, Yang Z, Chen YT, Ishiguro H, Uemura H, Kubota Y, et al. Promotion of bladder cancer development and progression by androgen receptor signals. J Natl Cancer Inst. 2007;99:558–68.

    Article  CAS  PubMed  Google Scholar 

  20. De Toni L, Guidolin D, De Filippis V, Tescari S, Strapazzon G, Santa Rocca M, et al. Osteocalcin and sex hormone binding globulin compete on a specific binding site of GPRC6A. Endocrinology. 2016;157:4473–86.

    Article  PubMed  Google Scholar 

  21. Pi M, Parrill AL, Quarles LD. GPRC6A mediates the non-genomic effects of steroids. J Biol Chem. 2010;285:39953–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Thomas P, Pang Y, Dong J, Berg AH. Identification and characterization of membrane androgen receptors in the ZIP9 zinc transporter subfamily: II. Role of human ZIP9 in testosterone-induced prostate and breast cancer cell apoptosis. Endocrinology. 2014;155:4250–65.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Thomas P, Pang Y, Dong J. Membrane androgen receptor characteristics of human ZIP9 (SLC39A) zinc transporter in prostate cancer cells: androgen-specific activation and involvement of an inhibitory G protein in zinc and MAP kinase signaling. Mol Cell Endocrinol. 2017;447:23–34.

    Article  CAS  PubMed  Google Scholar 

  24. Munnich N, Wernhart S, Hogstrand C, Schlomann U, Nimsky C, Bartsch JW. Expression of the zinc importer protein ZIP9/SLC39A9 in glioblastoma cells affects phosphorylation states of p53 and GSK-3beta and causes increased cell migration. Biometals. 2016;29:995–1004.

    Article  PubMed  Google Scholar 

  25. Thal DM, Glukhova A, Sexton PM, Christopoulos A. Structural insights into G-protein-coupled receptor allostery. Nature. 2018;559:45–53.

    Article  CAS  PubMed  Google Scholar 

  26. Tang X, Jin R, Qu G, Wang X, Li Z, Yuan Z, et al. GPR116, an adhesion G-protein-coupled receptor, promotes breast cancer metastasis via the Galphaq-p63RhoGEF-Rho GTPase pathway. Cancer Res. 2013;73:6206–18.

    Article  CAS  PubMed  Google Scholar 

  27. Gao ZG, Jacobson KA. On the selectivity of the Galphaq inhibitor UBO-QIC: a comparison with the Galphai inhibitor pertussis toxin. Biochem Pharm. 2016;107:59–66.

    Article  CAS  PubMed  Google Scholar 

  28. Berg AH, Rice CD, Rahman MS, Dong J, Thomas P. Identification and characterization of membrane androgen receptors in the ZIP9 zinc transporter subfamily: I. Discovery in female atlantic croaker and evidence ZIP9 mediates testosterone-induced apoptosis of ovarian follicle cells. Endocrinology. 2014;155:4237–49.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Niu Y, Altuwaijri S, Lai KP, Wu CT, Ricke WA, Messing EM, et al. Androgen receptor is a tumor suppressor and proliferator in prostate cancer. Proc Natl Acad Sci USA. 2008;105:12182–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Heinlein CA, Chang C. Androgen receptor in prostate cancer. Endocr Rev. 2004;25:276–308.

    Article  CAS  PubMed  Google Scholar 

  31. Rosner W, Hryb DJ, Khan MS, Nakhla AM, Romas NA. Androgen and estrogen signaling at the cell membrane via G-proteins and cyclic adenosine monophosphate. Steroids. 1999;64:100–6.

    Article  CAS  PubMed  Google Scholar 

  32. Lieberherr M, Grosse B. Androgens increase intracellular calcium concentration and inositol 1,4,5-trisphosphate and diacylglycerol formation via a pertussis toxin-sensitive G-protein. J Biol Chem. 1994;269:7217–23.

    CAS  PubMed  Google Scholar 

  33. Pi M, Quarles LD. GPRC6A regulates prostate cancer progression. Prostate. 2012;72:399–409.

    Article  CAS  PubMed  Google Scholar 

  34. Thomas P, Converse A, Berg HA. ZIP9, a novel membrane androgen receptor and zinc transporter protein. Gen Comp Endocrinol. 2018;257:130–6.

    Article  CAS  PubMed  Google Scholar 

  35. Izumi K, Taguri M, Miyamoto H, Hara Y, Kishida T, Chiba K, et al. Androgen deprivation therapy prevents bladder cancer recurrence. Oncotarget. 2014;5:12665–74.

    Article  PubMed  PubMed Central  Google Scholar 

  36. Shiota M, Yokomizo A, Takeuchi A, Imada K, Kiyoshima K, Inokuchi J, et al. Secondary bladder cancer after anticancer therapy for prostate cancer: reduced comorbidity after androgen-deprivation therapy. Oncotarget. 2015;6:14710–9.

    PubMed  PubMed Central  Google Scholar 

  37. Goldsmith ZG, Dhanasekaran DN. G protein regulation of MAPK networks. Oncogene. 2007;26:3122–42.

    Article  CAS  PubMed  Google Scholar 

  38. Wang JL, Yang MY, Xiao S, Sun B, Li YM, Yang LY. Downregulation of castor zinc finger 1 predicts poor prognosis and facilitates hepatocellular carcinoma progression via MAPK/ERK signaling. J Exp Clin Cancer Res. 2018;37:45.

    Article  PubMed  PubMed Central  Google Scholar 

  39. Huang B, Xiao E, Huang M. MEK/ERK pathway is positively involved in hypoxia-induced vasculogenic mimicry formation in hepatocellular carcinoma which is regulated negatively by protein kinase A. Med Oncol. 2015;32:408.

    Article  PubMed  Google Scholar 

  40. Liu Q, Zou R, Zhou R, Gong C, Wang Z, Cai T, et al. miR-155 regulates glioma cells invasion and chemosensitivity by p38 isforms in vitro. J Cell Biochem. 2015;116:1213–21.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Karen Wolf for help preparing the manuscript.

Funding

This work was supported by NIH grant (CA155477), in part by Taiwan Ministry of Health and Welfare Clinical Trial and Research Center of Excellence (MOHW104-TDU-B-212-113002), the National Natural Science Foundation of China (81572523), and the Hunan Province Funds for Distinguished Young Scientists of China (2016JJ1026).

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Authors and Affiliations

  1. Departments of Urology, Xiangya Hospital, Central South University, 410008, Changsha, China

    Jinbo Chen, Zhenyu Ou & Xiongbing Zu

  2. George Whipple Lab for Cancer Research, Departments of Pathology, Urology, Radiation Oncology, and The Wilmot Cancer Center, University of Rochester Medical Institute, Rochester, NY, USA

    Jinbo Chen, Fuju Chou, Shuyuan Yeh, Zhenyu Ou, Zhendong Xiang, Yin Sun, Edward Messing & Chawnshang Chang

  3. Sex Hormone Research Center, Departments of Urology and Medical Technology, China Medical University/Hospital, 404, Taichung, Taiwan

    Chihrong Shyr, Chiping Huang & Chawnshang Chang

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  1. Jinbo Chen
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Contributions

J.B.C., F.J.C., S.Y.Y., X.B.Z., and C.S.C. designed the experimental protocols. J.B.C., F.J.C., Z.Y.O., C.R.S., C.P.H., Z.D.X., and Y.S. performed the studies. J.B.C., F.J.C., S.Y.Y., and Z.D.X. analyzed the data. J.B.C., F.J.C., C.R.S., C.P.H., Y.S., E.M., and C.S.C. wrote the manuscript with contributions from all of the other authors.

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Correspondence to Xiongbing Zu or Chawnshang Chang.

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This study was approved by the Xiangya Hospital, Central South University Ethics Committee. Signed informed consents were obtained from all the patients. And the animal experiments were conducted strictly in accordance with the Animal Study Guidelines of University of Rochester Medical Center.

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Chen, J., Chou, F., Yeh, S. et al. Androgen dihydrotestosterone (DHT) promotes the bladder cancer nuclear AR-negative cell invasion via a newly identified membrane androgen receptor (mAR-SLC39A9)-mediated Gαi protein/MAPK/MMP9 intracellular signaling. Oncogene 39, 574–586 (2020). https://doi.org/10.1038/s41388-019-0964-6

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