Journal of Cancer Research and Clinical Oncology

, Volume 145, Issue 12, pp 2969–2982 | Cite as

Differential expression of p52 and RelB proteins in the metastatic and non-metastatic groups of uveal melanoma with patient outcome

  • Mithalesh Kumar Singh
  • Lata Singh
  • Kunzang Chosdol
  • Neelam Pushker
  • Neeru Saini
  • Rachna Meel
  • Sameer Bakhshi
  • Seema Sen
  • Seema KashyapEmail author
Original Article – Cancer Research



Non-canonical NFκB (NC-NFκB) pathway plays an influential role in metastasis, which promotes cancer proliferation and progression. The aim of the study was to examine the expression of NC-NFκB proteins and their correlation with clinicopathological factors associated with metastatic cases of uveal melanoma (UM) and with the patient outcome.


Expression of NC-NFκB proteins (p52, RelB, and co-expression of p52/RelB) was evaluated in 75 formalin-fixed cases of uveal melanoma by immunohistochemistry. Validation of nuclear immunoreactivity was done by western blotting. Transcriptional status of NC-NFκB genes was assessed in 60 fresh tumor tissues by quantitative real-time PCR. Co-immunoprecipitation was performed to determine the presence of native p52/RelB heterodimer in UM. Prognostic relevance was determined using Cox proportional hazard and Kaplan–Meier methods.


Immunohistochemical expression of p52, RelB, and their co-expression was observed in 81%, 68.7%, 56.2% of metastatic cases, respectively, while their expression was seen only in 38%, 33% and 30% of non-metastatic cases. Loss of BAP-1 was correlated with expression of p52 and RelB proteins. Co-immunoprecipitation assay confirmed the putative interaction of p52 with RelB protein in metastatic cases of uveal melanoma. Co-expression of p52/RelB and expression of p52 protein was significantly correlated with decreased metastasis-free survival (MFS) (p = 0.004; p = 0.002) and overall survival (OS) (p = 0.004; p = 0.032), while the RelB expression only correlated with reduced MFS (p = 0.003).


Our data showed that non-canonical NFκB proteins were significantly higher in metastatic cases and associated with poor outcome of the patients. Furthermore, the p52 protein could be used as a potential therapeutic biomarker for metastatic cases in uveal melanoma.


Uveal melanoma Metastasis p52 RelB Co-expression 



Mithalesh Kumar Singh is 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 immunohistochemistry.

Author contributions

All authors made substantial contributions to the study and they have approved the current version and agreed to publication.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Informed consent


Ethical approval

The study was conducted following the protocol approved by the institutional ethics committee, All India Institute of Medical Sciences (IESC/T-417/2015).


  1. AJCC Ophthalmic Oncology Task Force (2015) International validation of the American Joint Committee on Cancer’s 7th edition classification of uveal melanoma. JAMA Ophthalmol 133:376–383CrossRefGoogle Scholar
  2. Basseres DS, Baldwin AS (2006) Nuclear factor-kappaB and inhibitor of kappaB kinase pathways in oncogenic initiation and progression. Oncogene 25:6817–6830CrossRefGoogle Scholar
  3. Ben-Neriah Y, Karin M (2011) Inflammation meets cancer, with NF-kappaB as the matchmaker. Nat Immunol 12:715–723CrossRefGoogle Scholar
  4. Bronkhorst IH, Jager MJ (2012) Uveal melanoma: the inflammatory microenvironment. J Innate Immun 4:454–462CrossRefGoogle Scholar
  5. Cao Y, Bonizzi G, Seagroves TN, Greten FR, Johnson R et al (2001) IKKalpha provides an essential link between RANK signaling and cyclin D1 expression during mammary gland development. Cell 107:763–775CrossRefGoogle Scholar
  6. Demicco EG, Kavanagh KT, Romieu-Mourez R, Wang X, Shin SR et al (2005) RELB/p52 NF- B complexes rescue an early delay in mammary gland development in transgenic mice with targeted superrepressor I B-expression and promote carcinogenesis of the mammary gland. Mol Cell Biol 25:10136–10147CrossRefGoogle Scholar
  7. Dror R, Lederman M, Umezawa K et al (2010) Characterizing the involvement of the nuclear factor kappa B (NF kappa B) transcription factor in uveal melanoma. Investig Ophthalmol Vis Sci 51:1811–1816CrossRefGoogle Scholar
  8. Ehlers JP, Harbour JW (2006) Molecular pathobiology of uveal melanoma. Int Ophthalmol Clin 46:167–180CrossRefGoogle Scholar
  9. Gezgin G, Dogrusoz M, van Essen TH, Kroes WG, Luyten GP et al (2017) Genetic evolution of uveal melanoma guides the development of an inflammatory microenvironment. Cancer Immunol Immunother 66:903–912CrossRefGoogle Scholar
  10. Harbour JW, Onken MD, Roberson ED et al (2010) Frequent mutation of BAP1 in metastasizing uveal melanomas. Science 330:1410–1413CrossRefGoogle Scholar
  11. Kaliki S, Shields CL, Shields JA (2015) Uveal melanoma: estimating prognosis. Indian J Ophthalmol 63:93–102CrossRefGoogle Scholar
  12. Karin M, Cao Y, Greten FR, Li ZW (2002) NF-kappaB in cancer: from innocent bystander to major culprit. Nat Rev Cancer 2:301–310CrossRefGoogle Scholar
  13. Kath R, Hayungs J, Bornfeld N, Sauerwein W, Hoffken K, Seeber S (1993) Prognosis and treatment of disseminated uveal melanoma. Cancer 72:2219–2223CrossRefGoogle Scholar
  14. Koopmans AE, Verdijk RM, Brouwer RW et al (2014) Clinical significance of immunohistochemistry for detection of BAP1 mutations in uveal melanoma. Mod Pathol 27:1321–1330CrossRefGoogle Scholar
  15. Kujala E, Makitie T, Kivela T (2003) Very long-term prognosis of patients with malignant uveal melanoma. Investig Ophthalmol Vis Sci 44:4651–4659CrossRefGoogle Scholar
  16. Li Zhiqin, Han Chunfang, Feng Jing (2017) Relationship of the expression levels of XIAP and p53 genes in hepatocellular carcinoma and the prognosis of patients. Oncol Lett 14:4037–4042CrossRefGoogle Scholar
  17. Lorigan JG, Wallace S, Mavligit GM (1991) The prevalence and location of metastases from ocular melanoma: imaging study in 110 patients. AJR Am J Roentgenol 157:1279–1281CrossRefGoogle Scholar
  18. Lua J, Qayyum T, Edwards J, Roseweir AK (2018) The prognostic role of the non canonical nuclear factor-kappa B pathway in renal cell carcinoma patients. Urol Int 101:190–196CrossRefGoogle Scholar
  19. Maat W, Ly LV, Jordanova ES, de Wolff-Rouendaal D, Schalij-Delfos NE, Jager MJ (2008) Monosomy of chromosome 3 and an inflammatory phenotype occur together in uveal melanoma. Investig Ophthalmol Vis Sci 49:505–510CrossRefGoogle Scholar
  20. Mantovani A, Sica A (2005) Macrophages, innate immunity and cancer: balance, tolerance, and diversity. Curr Opin Immunol 22:231–237CrossRefGoogle Scholar
  21. Meir T, Dror R, Yu X, Qian J et al (2007) Molecular characteristics of liver metastases from uveal melanoma. Investig Ophthalmol Vis Sci 48:4890–4896CrossRefGoogle Scholar
  22. Miller LD, Smeds J, George J, Vega VB, Vergara L, Ploner A, Pawitan Y et al (2005) An expression signature for p53 status in human breast cancer predicts mutation status, transcriptional effects, and patient survival. Proc Natl Acad Sci USA 102:13550–13555CrossRefGoogle Scholar
  23. Mineva ND, Wang X, Yang S, Ying H et al (2009) Inhibition of RelB by 1,25-dihydroxyvitamin D3 promotes sensitivity of breast cancer cells to radiation. J Cell Physiol 220:593–599CrossRefGoogle Scholar
  24. Nguyen BT, Kim RS, Bretana ME et al (2018) Association between traditional clinical high-risk features and gene expression profile classification in uveal melanoma. Graefes Arch Clin Exp Ophthalmol 256:211–219CrossRefGoogle Scholar
  25. Oya M, Takayanagi A, Horiguchi A, Mizuno R, Ohtsubo M et al (2003) Increased nuclear factor-kB expression is related to the tumor development of renal cell carcinoma. Carcinogenesis 24:377–384CrossRefGoogle Scholar
  26. Prasad S, Ravindran J, Aggarwal BB (2010) NF-kappaB and cancer: how intimate is this relationship. Mol Cell Biochem 336(2010):25–37CrossRefGoogle Scholar
  27. Qin H, Zhou J, Zhou P, Xu J, Tang Z et al (2015) Prognostic significance of RelB over-expression in non-small cell lung cancer patients. Thorac Cancer 7:415–421CrossRefGoogle Scholar
  28. Rojo F, González-Pérez A, Furriol J, Nicolau MJ, Ferrer J, Burgués O et al (2016) Non-canonical NFκB pathway expression predicts outcome in borderline oestrogen receptor positive breast carcinoma. Br J Cancer 115:322–331CrossRefGoogle Scholar
  29. Saccani S, Pantano S, Natoli G (2003) Modulation of NF-kappaB activity by exchange of dimers. Mol Cell 11:1563–1574CrossRefGoogle Scholar
  30. Singh AD, Turell ME, Topham AK (2011) Uveal melanoma: trends in incidence, treatment, and survival. Ophthalmology 118:1881–1885CrossRefGoogle Scholar
  31. Ueda Y, Richmond A (2006) NFκB expression in melanoma. Pigment Cell Res 19:112–124CrossRefGoogle Scholar
  32. Wang B, Parobchak N, Rosen T (2012) RELB/NF-kappaB2 regulates corticotropin-releasing hormone in the human placenta. Mol Endocrinol 26:1356–1369CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 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.Department of BiosciencesJMINew DelhiIndia
  3. 3.Department of BiochemistryAIIMSNew DelhiIndia
  4. 4.Department of Ophthalmology, Dr. R. P. Centre for Ophthalmic SciencesAll India Institute of Medical SciencesNew DelhiIndia
  5. 5.Functional Genomics UnitInstitute of Genomics and Integrative BiologyNew DelhiIndia
  6. 6.Department of Medical OncologyIRCH, AIIMSNew DelhiIndia

Personalised recommendations