Loss of EGR3 is an independent risk factor for metastatic progression in prostate cancer

Abstract

Identification of pro-metastatic genomic alterations is urgently needed to help understand and prevent the fatal course of prostate cancer. Here, we found that the transcription factor EGR3, located at chromosome 8p21.3, is a critical metastasis suppressor. Aberrant deletion of EGR3 was found in up to 59.76% (deep deletions, 16.87%; shallow deletions, 42.89%) of prostate cancer patients. In informatics analysis, EGR3 loss was associated with prostate cancer progression and low survival rates. EGR3 expression inversely correlated with the expressions of epithelial-to-mesenchymal transition (EMT) and metastasis-related gene sets in prostate cancer tissues. In prostate cancer cells, EGR3 blocked the EMT process and suppressed cell migration and invasion. In a mouse model for cancer metastasis, EGR3 overexpression significantly suppressed bone metastases of PC3 and 22Rv1 prostate cancer cells. Mechanistically, EGR3 transcriptionally activated ZFP36, GADD45B, and SOCS3 genes by directly binding to their promoter regions. The EMT-inhibitory and tumor-suppressive roles of the EGR3 downstream genes were identified through in vitro and in silico analyses. Together, our results showed that EGR3 may be a biomarker to predict clinical outcomes and that it plays an important role in the metastatic progression of prostate cancer.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1: EGR3 deletion in prostate cancer patients.
Fig. 2: EGR3 protein levels are reduced in advanced prostate cancer and inversely correlate with patient survival.
Fig. 3: EGR3 expression negatively correlates with EMT and metastasis of prostate cancer.
Fig. 4: EGR3 inhibits EMT, migration, and invasion of prostate cancer cells.
Fig. 5: Transwell migration and invasion assays of prostate cancer cells.
Fig. 6: EGR3 represses bone metastasis of prostate cancer cells in vivo.
Fig. 7: EGR3 directly transactivates ZFP36, GADD45B, and SOCS3 genes to suppress EMT and cell migration.
Fig. 8: Clinical significance of EGR3 downstream target genes.

References

  1. 1.

    Brawley OW. Prostate cancer epidemiology in the United States. World J Urol. 2012;30:195–200.

    PubMed  Google Scholar 

  2. 2.

    Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin. 2019;69:7–34.

    Google Scholar 

  3. 3.

    Demichelis F, Attard G. A step toward functionally characterized prostate cancer molecular subtypes. Nat Med. 2013;19:966–7.

    CAS  PubMed  Google Scholar 

  4. 4.

    Reid AH, Attard G, Ambroisine L, Fisher G, Kovacs G, Brewer D, et al. Molecular characterisation of ERG, ETV1 and PTEN gene loci identifies patients at low and high risk of death from prostate cancer. Br J Cancer. 2010;102:678–84.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. 5.

    Kim SH, Kim SH, Joung JY, Lee GK, Hong EK, Kang KM, et al. Overexpression of ERG and Wild-Type PTEN are associated with favorable clinical prognosis and low biochemical recurrence in prostate cancer. PLoS ONE. 2015;10:e0122498.

    PubMed  PubMed Central  Google Scholar 

  6. 6.

    Kirby M, Hirst C, Crawford ED. Characterising the castration-resistant prostate cancer population: a systematic review. Int J Clin Pr. 2011;65:1180–92.

    CAS  Google Scholar 

  7. 7.

    Buttigliero C, Tucci M, Bertaglia V, Vignani F, Bironzo P, Di Maio M, et al. Understanding and overcoming the mechanisms of primary and acquired resistance to abiraterone and enzalutamide in castration resistant prostate cancer. Cancer Treat Rev. 2015;41:884–92.

    CAS  PubMed  Google Scholar 

  8. 8.

    Sartor O, de Bono JS. Metastatic prostate cancer. N Engl J Med. 2018;378:1653–4.

    PubMed  Google Scholar 

  9. 9.

    Parker C, Gillessen S, Heidenreich A, Horwich A, Committee EG. Cancer of the prostate: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2015;26(Suppl 5):v69–77.

    PubMed  Google Scholar 

  10. 10.

    Gomez-Martin D, Diaz-Zamudio M, Galindo-Campos M, Alcocer-Varela J. Early growth response transcription factors and the modulation of immune response: implications towards autoimmunity. Autoimmun Rev. 2010;9:454–8.

    CAS  PubMed  Google Scholar 

  11. 11.

    O’Donovan KJ, Levkovitz Y, Ahn D, Baraban JM. Functional comparison of Egr3 transcription factor isoforms: identification of an activation domain in the N-terminal segment absent from Egr3beta, a major isoform expressed in brain. J Neurochem. 2000;75:1352–7.

    PubMed  Google Scholar 

  12. 12.

    Decker EL, Nehmann N, Kampen E, Eibel H, Zipfel PF, Skerka C. Early growth response proteins (EGR) and nuclear factors of activated T cells (NFAT) form heterodimers and regulate proinflammatory cytokine gene expression. Nucleic Acids Res. 2003;31:911–21.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Tourtellotte WG, Milbrandt J. Sensory ataxia and muscle spindle agenesis in mice lacking the transcription factor Egr3. Nat Genet. 1998;20:87–91.

    CAS  PubMed  Google Scholar 

  14. 14.

    Eldredge LC, Gao XM, Quach DH, Li L, Han X, Lomasney J, et al. Abnormal sympathetic nervous system development and physiological dysautonomia in Egr3-deficient mice. Development 2008;135:2949–57.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. 15.

    Safford M, Collins S, Lutz MA, Allen A, Huang CT, Kowalski J, et al. Egr-2 and Egr-3 are negative regulators of T cell activation. Nat Immunol. 2005;6:472–80.

    CAS  PubMed  Google Scholar 

  16. 16.

    Liao F, Ji MY, Shen L, Qiu S, Guo XF, Dong WG. Decreased EGR3 expression is related to poor prognosis in patients with gastric cancer. J Mol Histol. 2013;44:463–8.

    CAS  PubMed  Google Scholar 

  17. 17.

    Suzuki T, Inoue A, Miki Y, Moriya T, Akahira J, Ishida T, et al. Early growth responsive gene 3 in human breast carcinoma: a regulator of estrogen-meditated invasion and a potent prognostic factor. Endocr Relat Cancer. 2007;14:279–92.

    CAS  PubMed  Google Scholar 

  18. 18.

    Vlietstra RJ, van Alewijk DC, Hermans KG, van Steenbrugge GJ, Trapman J. Frequent inactivation of PTEN in prostate cancer cell lines and xenografts. Cancer Res. 1998;58:2720–3.

    CAS  PubMed  Google Scholar 

  19. 19.

    Fraser M, Zhao H, Luoto KR, Lundin C, Coackley C, Chan N, et al. PTEN deletion in prostate cancer cells does not associate with loss of RAD51 function: implications for radiotherapy and chemotherapy. Clin Cancer Res. 2012;18:1015–27.

    CAS  PubMed  Google Scholar 

  20. 20.

    Logothetis CJ, Gallick GE, Maity SN, Kim J, Aparicio A, Efstathiou E, et al. Molecular classification of prostate cancer progression: foundation for marker-driven treatment of prostate cancer. Cancer Disco. 2013;3:849–61.

    CAS  Google Scholar 

  21. 21.

    Wang G, Zhao D, Spring DJ, DePinho RA. Genetics and biology of prostate cancer. Genes Dev. 2018;32:1105–40.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. 22.

    Gandhi J, Afridi A, Vatsia S, Joshi G, Joshi G, Kaplan SA, et al. The molecular biology of prostate cancer: current understanding and clinical implications. Prostate Cancer Prostatic Dis. 2018;21:22–36.

    CAS  PubMed  Google Scholar 

  23. 23.

    Jamaspishvili T, Berman DM, Ross AE, Scher HI, De Marzo AM, Squire JA, et al. Clinical implications of PTEN loss in prostate cancer. Nat Rev Urol. 2018;15:222–34.

    CAS  PubMed  Google Scholar 

  24. 24.

    Bova GS, Carter BS, Bussemakers MJ, Emi M, Fujiwara Y, Kyprianou N, et al. Homozygous deletion and frequent allelic loss of chromosome 8p22 loci in human prostate cancer. Cancer Res. 1993;53:3869–73.

    CAS  PubMed  Google Scholar 

  25. 25.

    El Gammal AT, Bruchmann M, Zustin J, Isbarn H, Hellwinkel OJ, Kollermann J, et al. Chromosome 8p deletions and 8q gains are associated with tumor progression and poor prognosis in prostate cancer. Clin Cancer Res. 2010;16:56–64.

    CAS  PubMed  Google Scholar 

  26. 26.

    Kluth M, Amschler NN, Galal R, Moller-Koop C, Barrow P, Tsourlakis MC, et al. Deletion of 8p is an independent prognostic parameter in prostate cancer. Oncotarget 2017;8:379–92.

    PubMed  Google Scholar 

  27. 27.

    Li S, Miao T, Sebastian M, Bhullar P, Ghaffari E, Liu M, et al. The transcription factors Egr2 and Egr3 are essential for the control of inflammation and antigen-induced proliferation of B and T cells. Immunity 2012;37:685–96.

    CAS  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Wendt MK, Balanis N, Carlin CR, Schiemann WP. STAT3 and epithelial-mesenchymal transitions in carcinomas. JAKSTAT. 2014;3:e28975.

    PubMed  PubMed Central  Google Scholar 

  29. 29.

    Brooks SA, Blackshear PJ. Tristetraprolin (TTP): interactions with mRNA and proteins, and current thoughts on mechanisms of action. Biochim Biophys Acta. 2013;1829:666–79.

    CAS  PubMed  PubMed Central  Google Scholar 

  30. 30.

    Lai WS, Carballo E, Strum JR, Kennington EA, Phillips RS, Blackshear PJ. Evidence that tristetraprolin binds to AU-rich elements and promotes the deadenylation and destabilization of tumor necrosis factor alpha mRNA. Mol Cell Biol. 1999;19:4311–23.

    CAS  PubMed  PubMed Central  Google Scholar 

  31. 31.

    Park JM, Lee TH, Kang TH. Roles of tristetraprolin in tumorigenesis. Int J Mol Sci. 2018;19:3384.

  32. 32.

    Yoon NA, Jo HG, Lee UH, Park JH, Yoon JE, Ryu J, et al. Tristetraprolin suppresses the EMT through the down-regulation of Twist1 and Snail1 in cancer cells. Oncotarget. 2016;7:8931–43.

    PubMed  PubMed Central  Google Scholar 

  33. 33.

    Michaelis KA, Knox AJ, Xu M, Kiseljak-Vassiliades K, Edwards MG, Geraci M, et al. Identification of growth arrest and DNA-damage-inducible gene beta (GADD45beta) as a novel tumor suppressor in pituitary gonadotrope tumors. Endocrinology. 2011;152:3603–13.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. 34.

    Ou DL, Shen YC, Yu SL, Chen KF, Yeh PY, Fan HH, et al. Induction of DNA damage-inducible gene GADD45beta contributes to sorafenib-induced apoptosis in hepatocellular carcinoma cells. Cancer Res. 2010;70:9309–18.

    CAS  PubMed  Google Scholar 

  35. 35.

    Huang H, Wang Q, Du T, Lin C, Lai Y, Zhu D, et al. Matrine inhibits the progression of prostate cancer by promoting expression of GADD45B. Prostate 2018;78:327–35.

    CAS  PubMed  Google Scholar 

  36. 36.

    Pio R, Jia Z, Baron VT, Mercola D. Early growth response 3 (Egr3) is highly over-expressed in non-relapsing prostate cancer but not in relapsing prostate cancer. PLoS ONE. 2013;8:e54096.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. 37.

    Baron VT, Pio R, Jia Z, Mercola D. Early growth response 3 regulates genes of inflammation and directly activates IL6 and IL8 expression in prostate cancer. Br J Cancer. 2015;112:755–64.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. 38.

    Adamson ED, Mercola D. Egr1 transcription factor: multiple roles in prostate tumor cell growth and survival. Tumour Biol. 2002;23:93–102.

    CAS  PubMed  Google Scholar 

  39. 39.

    Adamson E, de Belle I, Mittal S, Wang Y, Hayakawa J, Korkmaz K, et al. Egr1 signaling in prostate cancer. Cancer Biol Ther. 2003;2:617–22.

    CAS  PubMed  Google Scholar 

  40. 40.

    Gitenay D, Baron VT. Is EGR1 a potential target for prostate cancer therapy? Future Oncol. 2009;5:993–1003.

    CAS  PubMed  PubMed Central  Google Scholar 

  41. 41.

    Choi YJ, Kim I, Lee JE, Park JW. PIN1 transcript variant 2 acts as a long non-coding RNA that controls the HIF-1-driven hypoxic response. Sci Rep. 2019;9:10599.

    PubMed  PubMed Central  Google Scholar 

  42. 42.

    Taylor BS, Schultz N, Hieronymus H, Gopalan A, Xiao Y, Carver BS, et al. Integrative genomic profiling of human prostate cancer. Cancer Cell. 2010;18:11–22.

    CAS  PubMed  PubMed Central  Google Scholar 

  43. 43.

    Chandran UR, Ma C, Dhir R, Bisceglia M, Lyons-Weiler M, Liang W, et al. Gene expression profiles of prostate cancer reveal involvement of multiple molecular pathways in the metastatic process. BMC Cancer. 2007;7:64.

    PubMed  PubMed Central  Google Scholar 

  44. 44.

    Yu YP, Landsittel D, Jing L, Nelson J, Ren B, Liu L, et al. Gene expression alterations in prostate cancer predicting tumor aggression and preceding development of malignancy. J Clin Oncol. 2004;22:2790–9.

    CAS  PubMed  Google Scholar 

  45. 45.

    Varambally S, Yu J, Laxman B, Rhodes DR, Mehra R, Tomlins SA, et al. Integrative genomic and proteomic analysis of prostate cancer reveals signatures of metastatic progression. Cancer Cell. 2005;8:393–406.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors acknowledge the contribution of all the investigators at the participating study sites. This work was supported by the National Research Foundation of Korea (2019R1A2B5B03069677 and 2020R1A4A2002903).

Author information

Affiliations

Authors

Contributions

JWP and SHS designed the study. SHS, IK, JEL, and JWP wrote and revised the paper. SHS, IK, JEL, and ML performed cell-based experiments and analyzed biochemical data. SHS, IK, and JEL analyzed informatics.

Corresponding author

Correspondence to Jong-Wan Park.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Shin, S., Kim, I., Lee, J.E. et al. Loss of EGR3 is an independent risk factor for metastatic progression in prostate cancer. Oncogene 39, 5839–5854 (2020). https://doi.org/10.1038/s41388-020-01418-5

Download citation

Search