Journal of Cancer Research and Clinical Oncology

, Volume 144, Issue 9, pp 1665–1683 | Cite as

Association between pathologic factors and ERG expression in prostate cancer: finding pivotal networking

  • Seung-Ryeol Lee
  • Young-Deuk ChoiEmail author
  • Nam-Hoon ChoEmail author
Original Article – Cancer Research



To evaluate associations between pathologic factors and erythroblast transformation-specific (ETS)-related gene (ERG) expression in prostate cancer patients. Using next-generation sequencing, we identified target genes and regulatory networks.


ERG expression in 60 radical prostatectomies was compared with pathological findings by association rule mining with the Apriori algorithm. Whole-exome and RNA sequencing were performed on three formalin-fixed, paraffin-embedded ERG-positive and negative prostate cancer samples. A network diagram identifying dominant altered genes was constructed using Cytoscape open-source bioinformatics platform and GeneMania plugin.


Pathologic conditions positive for perineural invasion, apical margins, and Gleason score 3 + 4 = 7 were significantly more likely to be ERG-positive than other pathologic conditions (p = 0.0008), suggesting an association between ERG positivity, perineural invasion, apical margins, and Gleason score 3 + 4 = 7 (Firth’s logistic regression: OR 42.565, 95% CI 1.670–1084.847, p = 0.0232). Results of whole-exome and RNA sequencing identified 97 somatic mutations containing common mutated genes. Regulatory network analysis identified NOTCH1, MEF2C, STAT3, LCK, CACNA2D3, PCSK7, MEF2A, PDZD2, TAB1, and ASGR1 as pivotal genes. NOTCH1 appears to function as a hub, because it had the highest node degree and betweenness. NOTCH1 staining was found 8 of 60 specimens (13%), with a significant association between ERG and NOTCH1 positivity (p = 0.001).


Evaluating the association between ERG expression and pathologic factors, and identifying the regulatory network and pivotal hub may help to understand the clinical significance of ERG-positive prostate cancer.


Prostate cancer ETS (erythroblast transformation-specific) gene fusions TMPRSS2 (transmembrane protease serine 2 gene) ERG (ETS-related gene) Next-generation sequencing 



This study was supported by a grant from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Welfare, Republic of Korea (Grant number: HI13C2163) and by the Mid-Career Researcher Program through a National Research Foundation of Korea grant (No. 2016R1A2B4011115; CNH).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

This study was approved by the Institutional Review Board of Severance Hospital (4-2013-0857). All procedures performed in this study were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Written informed consent given by participants was exempted, because the study design was retrospective, and information was anonymized and de-identified prior to analysis.


  1. Artavanis Tsakonas S, Rand MD, Lake RJ (1999) Notch signaling: cell fate control and signal integration in development. Science 284:770–776. CrossRefPubMedGoogle Scholar
  2. Bethel CR, Faith D, Li X, Guan B, Hicks JL, Lan F, Jenkins RB, Bieberich CJ, De Marzo AM (2006) Decreased NKX3.1 protein expression in focal prostatic atrophy, prostatic intraepithelial neoplasia, and adenocarcinoma: association with gleason score and chromosome 8p deletion. Cancer Res 66:10683–10690. CrossRefPubMedGoogle Scholar
  3. Brossette SE, Sprague AP, Hardin JM, Waites KB, Jones WT, Moser SA (1998) Association rules and data mining in hospital infection control and public health surveillance. J Am Med Inform Assoc 5:373–381.
  4. Carvalho FL, Simons BW, Eberhart CG, Berman DM (2014) Notch signaling in prostate cancer: a moving target. Prostate 74:933–945. CrossRefPubMedPubMedCentralGoogle Scholar
  5. Chaux A, Albadine R, Toubaji A, Hicks J, Meeker A, Platz EA, De Marzo AM, Netto GJ (2011) Immunohistochemistry for ERG expression as a surrogate for TMPRSS2-ERG fusion detection in prostatic adenocarcinomas. Am J Surg Pathol 35:1014–1020. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Epstein JI, Egevad L, Amin MB, Delahunt B, Srigley JR, Humphrey PA (2016) The 2014 International Society of Urological Pathology (ISUP) consensus conference on Gleason grading of prostatic carcinoma: definition of grading patterns and proposal for a new grading system. Am J Surg Pathol 40:244–252.
  7. Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J, Hornik K, Hothorn T, Huber W, Iacus S, Irizarry R, Leisch F, Li C, Maechler M, Rossini AJ, Sawitzki G, Smith C, Smyth G, Tierney L, Yang JY, Zhang J (2004) Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 5:R80-R80. CrossRefGoogle Scholar
  8. Hedegaard J, Thorsen K, Lund MK, Hein AK, Hamilton Dutoit SJ, Vang S, Nordentoft I, Birkenkamp Demtröder K, Kruhøffer M, Hager H, Knudsen B, Andersen CL, Sørensen KD, Pedersen JS, Ørntoft TF, Dyrskjøt L (2014) Next-generation sequencing of RNA and DNA isolated from paired fresh-frozen and formalin-fixed paraffin-embedded samples of human cancer and normal tissue. PLoS One 9:e98187. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Kimura T, Furusato B, Miki J, Yamamoto T, Hayashi N, Takahashi H, Kamata Y, van Leenders GJ, Visakorpi T, Egawa S (2012) Expression of ERG oncoprotein is associated with a less aggressive tumor phenotype in Japanese prostate cancer patients. Pathol Int 62:742–748. CrossRefPubMedGoogle Scholar
  10. Krstanoski Z, Vokac NK, Zagorac A, Pospihalj B, Munda M, Dzeroski S, Golouh R (2016) TMPRSS2:ERG gene aberrations may provide insight into pT stage in prostate cancer. BMC Urol 16:35. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Langmead B, Salzberg SL (2012) Fast gapped-read alignment with Bowtie 2. Nat Methods 9:357–359. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Lefort K, Ostano P, Mello-Grand M, Calpini V, Scatolini M, Farsetti A, Dotto GP, Chiorino G (2016) Dual tumor suppressing and promoting function of Notch1 signaling in human prostate cancer. Oncotarget 7:48011–48026. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Lu LI, Zhang H, Pang J, Hou GL, Lu MH, Gao X (2016) ERG rearrangement as a novel marker for predicting the extra-prostatic extension of clinically localised prostate cancer. Oncol Lett 11:2532–2538. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Manson Bahr D, Ball R, Gundem G, Sethia K, Mills R, Rochester M, Goody V, Anderson E, O’Meara S, Flather M, Keeling M, Yazbek Hanna M, Hurst R, Curley H, Clark J, Brewer DS, McDermott U, Cooper C (2015) Mutation detection in formalin-fixed prostate cancer biopsies taken at the time of diagnosis using next-generation DNA sequencing. J Clin Pathol 68:212–217. CrossRefPubMedGoogle Scholar
  15. Mohamed AA, Tan SH, Xavier CP, Katta S, Huang W, Ravindranath L, Jamal M, Li H, Srivastava M, Srivatsan ES, Sreenath TL, McLeod DG, Srinivasan A, Petrovics G, Dobi A, Srivastava S (2017) Synergistic activity with NOTCH inhibition and androgen ablation in ERG-positive prostate cancer cells. Mol Cancer Res 15:1308–1317. CrossRefPubMedGoogle Scholar
  16. Montojo J, Zuberi K, Rodriguez H, Kazi F, Wright G, Donaldson SL, Morris Q, Bader GD (2010) GeneMANIA Cytoscape plugin: fast gene function predictions on the desktop. Bioinformatics 26:2927–2928. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Neil JR, Schiemann WP (2008) Altered TAB1:I kappaB kinase interaction promotes transforming growth factor beta-mediated nuclear factor-kappaB activation during breast cancer progression. Cancer Res 68:1462–1470. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Park K, Tomlins SA, Mudaliar KM, Chiu YL, Esgueva R, Mehra R, Suleman K, Varambally S, Brenner JC, MacDonald T, Srivastava A, Tewari AK, Sathyanarayana U, Nagy D, Pestano G, Kunju LP, Demichelis F, Chinnaiyan AM, Rubin MA (2010) Antibody-based detection of ERG rearrangement-positive prostate cancer. Neoplasia 12:590–598CrossRefPubMedPubMedCentralGoogle Scholar
  19. Perner S, Mosquera J, Demichelis F, Hofer MD, Paris PL, Simko J, Collins C, Bismar TA, Chinnaiyan AM, De Marzo AM, Rubin MA (2007) TMPRSS2-ERG fusion prostate cancer: an early molecular event associated with invasion. Am J Surg Pathol 31:882–888. CrossRefPubMedGoogle Scholar
  20. Pettersson A, Graff RE, Bauer SR, Pitt MJ, Lis RT, Stack EC, Martin NE, Kunz L, Penney KL, Ligon AH, Suppan C, Flavin R, Sesso HD, Rider JR, Sweeney C, Stampfer MJ, Fiorentino M, Kantoff PW, Sanda MG, Giovannucci EL, Ding EL, Loda M, Mucci LA (2012) The TMPRSS2:ERG rearrangement, ERG expression, and prostate cancer outcomes: a cohort study and meta-analysis. Cancer Epidemiol Biomarkers Prev 21:1497–1509. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Scardoni G, Petterlini M, Laudanna C (2009) Analyzing biological network parameters with CentiScaPe. Bioinformatics 25:2857–2859. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Sethi S, Macoska J, Chen W, Sarkar FH (2010) Molecular signature of epithelial-mesenchymal transition (EMT) in human prostate cancer bone metastasis. Am J Transl Res 3:90–99PubMedPubMedCentralGoogle Scholar
  23. Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, Amin N, Schwikowski B, Ideker T (2003) Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res 13:2498–2504. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Shin SJ, Park CK, Park SY, Jang WS, Lee JY, Choi YD, Cho NH (2016) Total intraglandular and index tumor volumes predict biochemical recurrence in prostate cancer. Virchows Arch 469:305–312. CrossRefPubMedGoogle Scholar
  25. Smoot ME, Ono K, Ruscheinski J, Wang P, Ideker T (2011) Cytoscape 2.8: new features for data integration and network visualization. Bioinformatics 27:431–432. CrossRefPubMedGoogle Scholar
  26. Tomlins SA, Rhodes DR, Perner S, Dhanasekaran SM, Mehra R, Sun X, Varambally S, Cao X, Tchinda J, Kuefer R, Lee C, Montie JE, Shah RB, Pienta KJ, Rubin MA, Chinnaiyan AM (2005) Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science 310:644–648. CrossRefPubMedGoogle Scholar
  27. Tomlins SA, Bjartell A, Chinnaiyan AM, Jenster G, Nam RK, Rubin MA, Schalken JA (2009) ETS gene fusions in prostate cancer: from discovery to daily clinical practice. Eur Urol 56:275–286. CrossRefPubMedGoogle Scholar
  28. Vargas JE, Porto BN, Puga R, Stein RT, Pitrez PM (2016) Identifying a biomarker network for corticosteroid resistance in asthma from bronchoalveolar lavage samples. Mol Biol Rep 43:697–710. CrossRefPubMedGoogle Scholar
  29. Wang X, Leow CC, Zha J, Tang Z, Modrusan Z, Radtke F, Aguet M, de Sauvage FJ, Gao W (2006) Notch signaling is required for normal prostatic epithelial cell proliferation and differentiation. Dev Biol 290:66–80. CrossRefPubMedGoogle Scholar
  30. Wright A, Chen ES, Maloney FL (2010) An automated technique for identifying associations between medications, laboratory results and problems. J Biomed Inform 43:891–901. CrossRefPubMedGoogle Scholar
  31. Xu B, Chevarie-Davis M, Chevalier S, Scarlata E, Zeizafoun N, Dragomir A, Tanguay S, Kassouf W, Aprikian A, Brimo F (2014) The prognostic role of ERG immunopositivity in prostatic acinar adenocarcinoma: a study including 454 cases and review of the literature. Hum Pathol 45:488–497. CrossRefPubMedGoogle Scholar
  32. Zhu H, Zhou X, Redfield S, Lewin J, Miele L (2013) Elevated Jagged-1 and Notch-1 expression in high grade and metastatic prostate cancers. Am J Transl Res 5:368–378PubMedPubMedCentralGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of Urology, CHA Bundang Medical CenterCHA University College of MedicineSeongnamSouth Korea
  2. 2.Department of UrologyYonsei University College of MedicineSeoulSouth Korea
  3. 3.Department of PathologyYonsei University College of MedicineSeoulSouth Korea

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