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Constitutive expression of c-REL in uveal melanoma patients: correlation with clinicopathological parameters and patient outcome

  • M. K. Singh
  • L. Singh
  • N. Pushker
  • K. Chosdol
  • S. Bakhshi
  • R. Meel
  • S. Sen
  • S. KashyapEmail author
Research Article
  • 20 Downloads

Abstract

Purpose

Uveal melanoma (UM) is the most common intraocular cancer with a high mortality rate that requires new research in the field of prevention and treatment. c-REL is a member of the nuclear factor κB (NF-κB) transcription factor family and an emerging regulator of tumorigenesis. Therefore, the objective of the study is to evaluate the constitutive expression of c-REL in uveal melanoma patients and its prognostic significance.

Methods

Detection of c-REL expression was carried out by immunohistochemistry in all 75 patients, and qRT-PCR performed on 58 fresh cases of uveal melanoma along with IL-6 status. Immunoblot was performed to validate immunohistochemistry results. Expression of c-REL protein correlated with clinicopathological parameters and overall survival of patients.

Results

Immunohistochemistry results revealed nuclear expression of the c-REL protein (56%) in our cases. Out of 75 cases, 31 cases showed nuclear expression, and 11 cases had cytoplasmic expression. qRT-PCR showed upregulation of the REL gene in 56.89% cases at the transcriptional level. There was a statistically significant difference in the overall survival of patients with c-REL nuclear immunopositivity (p = 0.0048). On multivariate analysis, scleral invasion and c-REL nuclear expression found to be an independent prognostic factor (p < 0.05)

Conclusions

To the best of our knowledge, this was the first study reporting the expression of the c-REL protein in uveal melanoma. Strong nuclear immunoexpression of c-Rel suggests NFκB pathway activation which might be involved in the progression of the disease. Differential expression of c-REL protein may be used as an attractive target for the development of anticancer strategies.

Keywords

Uveal melanoma c-REL Overall survival 

Notes

Acknowledgements

Mithalesh Kumar Singh was thankful to Indian Council of Medical Research for providing Senior Research Fellowship (SRF). We are grateful to Mr. Pankaj Kumar for providing technical support in IHC.

Author contribution

MKS and SK are responsible for the conception or design of the work. MKS, LS and KC contribute the acquisition, analysis, or interpretation of data for the work. NP, and RM provide the tissue samples. SB helps in the follow-up of the patients. SS helps in reviewing the histopathology slides. All authors finally approved the manuscript version to be published. SK is the guarantor of the article.

Compliance with ethical standards:

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The study was approved by Institutional Ethical Committee, All India Institute of Medical Sciences, New Delhi, India and conducted in accordance with the ethical standards.

Informerd consent

Yes.

Supplementary material

12094_2019_2247_MOESM1_ESM.tif (89.3 mb)
Supplementary material 1 (Tif 91461 kb)

References

  1. 1.
    Amaro A, Gangemi R, Piaggio F, et al. The biology of uveal melanoma. Cancer Metastasis Rev. 2017;36:109–40.CrossRefGoogle Scholar
  2. 2.
    Nathan P, Cohen V, Coupland S, et al. United Kingdom Uveal Melanoma Guideline Development Working Group. Eur J Cancer. 2015;51:2404–12.CrossRefGoogle Scholar
  3. 3.
    Diener-West M, Reynolds SM, Agugliaro DJ, et al. Collaborative Ocular Melanoma Study Group Development of metastatic disease after enrollment in the COMS trials for treatment of choroidal melanoma: collaborative ocular melanoma study group report no. 26. Arch Ophthalmol. 2005;123: 1639–1643.Google Scholar
  4. 4.
    Rietschel P, Panageas KS, Hanlon C, et al. Variates of survival in metastatic uveal melanoma. J Clin Oncol. 2005;23:7576.CrossRefGoogle Scholar
  5. 5.
    Damato B. Does ocular treatment of uveal melanoma influence survival? Br J Cancer. 2010;103:285–90.CrossRefGoogle Scholar
  6. 6.
    Li Q, Verma IM. NF-κB regulation in the immune system. Nat Rev Immunol. 2002;2:725–34.CrossRefGoogle Scholar
  7. 7.
    Karin M, Cao Y, Greten FR, Li ZW. NF-kB in cancer: from innocent bystander to major culprit. Nat Rev Cancer. 2002;2:301–10.CrossRefGoogle Scholar
  8. 8.
    Ben-Neriah Y, Karin M. Inflammation meets cancer, with NFkappaB as the matchmaker. Nat Immunol. 2011;12:715–23.CrossRefGoogle Scholar
  9. 9.
    Grivennikov SI, Karin M. Dangerous liaisons: STAT3 and NF kappa B collaboration and crosstalk in cancer. Cytokine Growth Factor Rev. 2010;21:11–9.CrossRefGoogle Scholar
  10. 10.
    Gilmore TD, Gerondakis S. The c-REL transcription factor in development and disease. Genes Cancer. 2011;2:695–711.CrossRefGoogle Scholar
  11. 11.
    Wang Y, Rickman BH, Poutahidis T, et al. c-REL is essential for the development of innate and T cell-induced colitis. J Immunol. 2008;175:8118–255.CrossRefGoogle Scholar
  12. 12.
    Compagno M, Lim WK, Grunn A, et al. Mutations of multiple genes cause deregulation of NF-kB in diffuse large B-cell lymphoma. Nature. 2009;459:717–21.CrossRefGoogle Scholar
  13. 13.
    Hamoudi RA, Appert A, Ye H, et al. Differential expression of NF-kB target genes in MALT lymphoma with and without chromosome translocation: insights into molecular mechanism. Leukemia. 2010;24:1487–97.CrossRefGoogle Scholar
  14. 14.
    Geismann C, Grohmann F, Sebens S, et al. c-REL is a critical mediator of NF-kB dependent TRAIL resistance of pancreatic cancer cells. Cell Death Dis. 2014;5:e1455.CrossRefGoogle Scholar
  15. 15.
    Cogswell PC, Guttridge DC, Funkhouser WK, Baldwin AS Jr. Selective activation of NF-kB subunits in human breast cancer: potential roles for NF-kB2/p52 and for Bcl-3. Oncogene. 2000;19:1123–31.CrossRefGoogle Scholar
  16. 16.
    Gieling RG, Elsharkawy AM, Caamano JH, et al. The c-REL subunit of nuclear factor-kB regulates murine liver inflammation, wound-healing, and hepatocyte proliferation. Hepatology. 2010;51:922–31.CrossRefGoogle Scholar
  17. 17.
    Elsharkawy AM, Mann DA. Nuclear factor-kB and the hepatic inflammation-fibrosis-cancer axis. Hepatology. 2007;46:590–7.CrossRefGoogle Scholar
  18. 18.
    McNulty SE, del Rosario R, Cen D, Meyskens FL Jr, Yang S. Comparative expression of NFkappa B proteins in melanocytes of normal skin vs. benign intradermal naevus and human metastatic melanoma biopsies. Pigment Cell Res. 2004;17:173–180.CrossRefGoogle Scholar
  19. 19.
    Wang Y, Rickman BH, Poutahidis T, et al. c-REL is essential for the development of innate and T cell-induced colitis. J Immunol. 2008;180:8118–255.CrossRefGoogle Scholar
  20. 20.
    Gilmore TD, Gerondakis S. The c-REL transcription factor in development and disease. Genes Cancer. 2011;2:695–711.CrossRefGoogle Scholar
  21. 21.
    Lu YC, Kim I, Lye E, Shen F, Suzuki N, Suzuki S, Gerondakis S, Akira S, Gaffen SL, Yeh WC, Ohashi PS. Differential role for c-Rel and C/EBPbeta/delta in TLR-mediated induction of proinflammatory cytokines. J Immunol. 2009;182:7212–21.CrossRefGoogle Scholar
  22. 22.
    Dror R, Lederman M, Umezawa K, Barak V, Pe'er J, Chowers I. Characterizing the involvement of the nuclear factor-kappa B (NFκB) transcription factor in uveal melanoma. Invest Ophthalmol Vis Sci. 2010;51:1811–6.CrossRefGoogle Scholar
  23. 23.
    Liu N, Sun Q, Chen J, Li J, Zeng Y, Zhai S, Li P, Wang B, Wang X. MicroRNA-9 suppresses uveal melanoma cell migration and invasion through the NF-κB1 pathway. Oncol Rep. 2012;28:961–8.PubMedGoogle Scholar
  24. 24.
    Makitie T, Summanen P, Tarkkanen A, Kivelä T. Tumor-infiltrating macrophages (CD68(+) cells) and prognosis in malignant uveal melanoma. Invest Ophthalmol Vis Sci. 2001;42(7):1414–21.PubMedGoogle Scholar
  25. 25.
    McLean IW, Foster WD, Zimmerman LE, Gamel JW. Modifications of Callender’s classification of uveal melanoma at the Armed Forces Institute of Pathology. Am J Ophthalmol. 1983;96:502–9.CrossRefGoogle Scholar
  26. 26.
    The AJCC Ophthalmic Oncology Task Force. International Validation of the American Joint Committee on Cancer’s 7th Edition Classification of Uveal Melanoma. JAMA Ophthalmol. 2015; 133: 376–383.CrossRefGoogle Scholar
  27. 27.
    Li Z, Han C, Feng J. Relationship of the expression levels of XIAP and p53 genes in hepatocellular carcinoma and the prognosis of patients. Oncol Lett. 2017;14(4):4037–42.CrossRefGoogle Scholar
  28. 28.
    Rodig SJ, Savage KJ, LaCasce AS, et al. Expression of TRAF1 and nuclear c-REL distinguishes primary mediastinal large cell lymphoma from other types of diffuse large B-cell lymphoma. Am J Surg Pathol. 2007;31:106–12.CrossRefGoogle Scholar
  29. 29.
    Tarlan B, Kıratlı H. Uveal melanoma: current trends in diagnosis and management. Turk J Ophthalmol. 2016;46:123–37.CrossRefGoogle Scholar
  30. 30.
    Shields CL, Shields JA, Kiratli H, De Potter P, Cater JR. Risk factors for growth and metastasis of small choroidal melanocytic lesions. Ophthalmology. 1995;102:1351–61.CrossRefGoogle Scholar
  31. 31.
    Shields CL, Furuta M, Thangappan A, et al. Metastasis of uveal melanoma millimeter-by-millimeter in 8033 consecutive eyes. Arch Ophthalmol. 2009;127:989–98.CrossRefGoogle Scholar
  32. 32.
    Diener-West M, Reynolds SM, Agugliaro DJ, et al. Collaborative Ocular Melanoma Study Group Development of metastatic disease after enrollment in the COMS trials for treatment of choroidal melanoma: collaborative ocular melanoma study group report no. 26. Arch Ophthalmol. 2005; 123: 1639–1643.Google Scholar
  33. 33.
    Singh AD, Shields CL, Shields JA. Prognostic factors in uveal melanoma. Melanoma Res. 2001;11:255–63.CrossRefGoogle Scholar
  34. 34.
    Jager MJ, Ly LV, El Filali M, et al. Macrophages in uveal melanoma and in experimental ocular tumor models: Friends or foes? Prog Retin Eye Res. 2011;30:129–46.CrossRefGoogle Scholar
  35. 35.
    Liu T, Zhang L, Joo D, Sun SC. NF-κB signaling in inflammation. Signal Transduct Target Ther. 2017; 2:17023.Google Scholar
  36. 36.
    Terai M, Mastrangleo MJ, Sato T. Immunological aspect of the liver and metastatic uveal melanoma. J Cancer Metastasis Treat. 2017;3:231–43.CrossRefGoogle Scholar
  37. 37.
    Weniger MA, Gesk S, Ehrlich S, et al. Gains of REL in primary mediastinal B-cell lymphoma coincide with nuclear accumulation of REL protein. Gene Chromosome Cancer. 2007;46:406–15.CrossRefGoogle Scholar
  38. 38.
    Li L, Xu-Monette ZY, Ok CY, Tzankov A, Manyam GC, Sun R, Visco C, Zhang M, et al. Prognostic impact of c-REL nuclear expression and REL amplification and crosstalk between c-REL and the p53 pathway in diffuse large B-cell lymphoma. Oncotarget. 2015;15:23517–80.Google Scholar
  39. 39.
    Barth TF, Martin-Subero JI, Joos S, et al. Gains of 2p involving the REL locus correlate with nuclear c-REL protein accumulation in neoplastic cells of classical Hodgkin lymphoma. Blood. 2003;101:3681–6.CrossRefGoogle Scholar
  40. 40.
    Curry CV, Ewton AA, Olsen RJ, et al. Prognostic impact of c-REL expression in diffuse large B-cell lymphoma. J Hematop. 2009;2:20–6.CrossRefGoogle Scholar

Copyright information

© Federación de Sociedades Españolas de Oncología (FESEO) 2019

Authors and Affiliations

  1. 1.Department of Ocular Pathology, Dr. R. P. Centre for Ophthalmic SciencesAll India Institute of Medical SciencesNew DelhiIndia
  2. 2.Departrment of BiosciencesJMINew DelhiIndia
  3. 3.Department of Ophthalmology, Dr. R. P. Centre for Ophthalmic SciencesAll India Institute of Medical SciencesNew DelhiIndia
  4. 4.Department of BiochemistryAIIMSNew DelhiIndia
  5. 5.Department of Medical OncologyIRCH, AIIMSNew DelhiIndia

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