Alternative NF-κB signaling promotes colorectal tumorigenesis through transcriptionally upregulating Bcl-3


Multiple studies have shown that chronic inflammation is closely related to the occurrence and development of colorectal cancer (CRC). Classical NF-κB signaling, the key factor in controlling inflammation, has been found to be of great importance to CRC development. However, the role of alternative NF-κB signaling in CRC is still elusive. Here, we found aberrant constitutive activation of alternative NF-κB signaling both in CRC tissue and CRC cells. Knockdown of RelB downregulates c-Myc and upregulates p27Kip1 protein level, which inhibits CRC cell proliferation and retards CRC xenograft growth. Conversely, overexpression of RelB increases proliferation of CRC cells. In addition, we revealed a significant correlation between Bcl-3 and RelB in CRC tissues. The expression of RelB was consistent with the expression of Bcl-3 and the phosphorylation of Bcl-3 downstream proteins p-Akt (S473) and p-GSK3β (S9). Bcl-3 overexpression can restore the phenotype changes caused by RelB knockdown. Importantly, we demonstrated that alternative NF-κB transcriptional factor (p52:RelB) can directly bind to the promoter region of Bcl-3 gene and upregulate its transcription. Moreover, the expression of RelB, NF-κB2 p52, and Bcl-3 was associated with poor survival of CRC patients. Taken together, these results represent that alternative NF-κB signaling may function as an oncogenic driver in CRC, and also provide new ideas and research directions for the pathogenesis, prevention, and treatment of other inflammatory-related diseases.

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  1. 1.

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

    Article  Google Scholar 

  2. 2.

    Siegel RL, Miller KD, Fedewa SA, Ahnen DJ, Meester RGS, Barzi A, et al. Colorectal cancer statistics, 2017. CA Cancer J Clin. 2017;67:177–93.

    Article  Google Scholar 

  3. 3.

    Karin M, NF-kB and cancer: mechanisms and targets. Mol Carcinog. 2006;45:355–61.

    CAS  Article  Google Scholar 

  4. 4.

    Ghosh MSHS. Shared principles in NF-κB signaling. Cell. 2008;132:344–62.

    Article  Google Scholar 

  5. 5.

    Terzic J, Grivennikov S, Karin E, Karin M. Inflammation and colon cancer. Gastroenterology. 2010;138:2101–14 e5.

    CAS  Article  Google Scholar 

  6. 6.

    Sakamoto K, Maeda S, Hikiba Y, Nakagawa H, Hayakawa Y, Shibata W, et al. Constitutive NF-kappaB activation in colorectal carcinoma plays a key role in angiogenesis, promoting tumor growth. Clin Cancer Res. 2009;15:2248–58.

    CAS  Article  Google Scholar 

  7. 7.

    Allen IC, Wilson JE, Schneider M, Lich JD, Roberts RA, Arthur JC, et al. NLRP12 suppresses colon inflammation and tumorigenesis through the negative regulation of noncanonical NF-kappaB signaling. Immunity. 2012;36:742–54.

    CAS  Article  PubMed Central  Google Scholar 

  8. 8.

    Dejardin E, Deregowski V, Chapelier M, Jacobs N, Gielen J, Merville MP, et al. Regulation of NF-kappaB activity by I kappaB-related proteins in adenocarcinoma cells. Oncogene. 1999;18:2567–77.

    CAS  Article  Google Scholar 

  9. 9.

    Lessard L, Begin LR, Gleave ME, Mes-Masson AM, Saad F. Nuclear localisation of nuclear factor-kappaB transcription factors in prostate cancer: an immunohistochemical study. Br J Cancer. 2005;93:1019–23.

    CAS  Article  PubMed Central  Google Scholar 

  10. 10.

    Annunziata CM, Davis RE, Demchenko Y, Bellamy W, Gabrea A, Zhan F, et al. Frequent engagement of the classical and alternative NF-kappaB pathways by diverse genetic abnormalities in multiple myeloma. Cancer Cell. 2007;12:115–30.

    CAS  Article  PubMed Central  Google Scholar 

  11. 11.

    Wharry CE, Haines KM, Carroll RG, May MJ. Constitutive non-canonical NFkappaB signaling in pancreatic cancer cells. Cancer Biol Ther. 2009;8:1567–76.

    CAS  Article  PubMed Central  Google Scholar 

  12. 12.

    Hailfinger S, Nogai H, Pelzer C, Jaworski M, Cabalzar K, Charton JE, et al. Malt1-dependent RelB cleavage promotes canonical NF-kappaB activation in lymphocytes and lymphoma cell lines. Proc Natl Acad Sci USA. 2011;108:14596–601.

    CAS  Article  Google Scholar 

  13. 13.

    Jacque E, Billot K, Authier H, Bordereaux D, Baud V. RelB inhibits cell proliferation and tumor growth through p53 transcriptional activation. Oncogene. 2013;32:2661–9.

    CAS  Article  Google Scholar 

  14. 14.

    Xu Y, Josson S, Fang F, Oberley TD, St Clair DK, Wan XS, et al. RelB enhances prostate cancer growth: implications for the role of the nuclear factor-kappaB alternative pathway in tumorigenicity. Cancer Res. 2009;69:3267–71.

    CAS  Article  PubMed Central  Google Scholar 

  15. 15.

    Nottingham LK, Yan CH, Yang X, Si H, Coupar J, Bian Y, et al. Aberrant IKKalpha and IKKbeta cooperatively activate NF-kappaB and induce EGFR/AP1 signaling to promote survival and migration of head and neck cancer. Oncogene. 2014;33:1135–47.

    CAS  Article  Google Scholar 

  16. 16.

    Wang X, Belguise K, Kersual N, Kirsch KH, Mineva ND, Galtier F, et al. Oestrogen signalling inhibits invasive phenotype by repressing RelB and its target BCL2. Nat Cell Biol. 2007;9:470–8.

    CAS  Article  PubMed Central  Google Scholar 

  17. 17.

    Ge QL, Liu SH, Ai ZH, Tao MF, Ma L, Wen SY, et al. RelB/NF-kappaB links cell cycle transition and apoptosis to endometrioid adenocarcinoma tumorigenesis. Cell Death Dis. 2016;7:e2402.

    CAS  Article  PubMed Central  Google Scholar 

  18. 18.

    Vallabhapurapu SD, Noothi SK, Pullum DA, Lawrie CH, Pallapati R, Potluri V, et al. Transcriptional repression by the HDAC4-RelB-p52 complex regulates multiple myeloma survival and growth. Nat Commun. 2015;6:8428.

    CAS  Article  Google Scholar 

  19. 19.

    Ohno H, Takimoto G, McKeithan TW. The candidate proto-oncogene bcl-3 is related to genes implicated in cell lineage determination and cell cycle control. Cell. 1990;60:991–7.

    CAS  Article  Google Scholar 

  20. 20.

    Bours V, Villalobos J, Burd PR, Kelly K, Siebenlist U. Cloning of a mitogen-inducible gene encoding a kappa B DNA-binding protein with homology to the rel oncogene and to cell-cycle motifs. Nature. 1990;348:76–80.

    CAS  Article  PubMed Central  Google Scholar 

  21. 21.

    Tanaka S, Nishigaki H, Nakagawa H, Okuda T, Nishida K, Tsuda S, et al. Reciprocal t(14;19)(q32.3; q13.1) in a patient with B-cell lymphoma. Cancer Genet Cytogenet. 1990;49:219–24.

    CAS  Article  PubMed Central  Google Scholar 

  22. 22.

    Naumann M, Wulczyn FG, Scheidereit C. The NF-kappa B precursor p105 and the proto-oncogene product Bcl-3 are I kappa B molecules and control nuclear translocation of NF-kappa B. EMBO J. 1993;12:213–22.

    CAS  Article  PubMed Central  Google Scholar 

  23. 23.

    Nolan GP, Fujita T, Bhatia K, Huppi C, Liou HC, Scott ML, et al. The bcl-3 proto-oncogene encodes a nuclear I kappa B-like molecule that preferentially interacts with NF-kappa B p50 and p52 in a phosphorylation-dependent manner. Mol Cell Biol. 1993;13:3557–66.

    CAS  Article  PubMed Central  Google Scholar 

  24. 24.

    Bours V, Franzoso G, Azarenko V, Park S, Kanno T, Brown K, et al. The oncoprotein Bcl-3 directly transactivates through kappa B motifs via association with DNA-binding p50B homodimers. Cell. 1993;72:729–39.

    CAS  Article  Google Scholar 

  25. 25.

    Maldonado V, Melendez-Zajgla J. Role of Bcl-3 in solid tumors. Mol Cancer. 2011;10:152.

    CAS  Article  PubMed Central  Google Scholar 

  26. 26.

    Liu Z, Jiang Y, Hou Y, Hu Y, Cao X, Tao Y, et al. The IkappaB family member Bcl-3 stabilizes c-Myc in colorectal cancer. J Mol Cell Biol. 2013;5:280–2.

    CAS  Article  PubMed Central  Google Scholar 

  27. 27.

    Urban BC, Collard TJ, Eagle CJ, Southern SL, Greenhough A, Hamdollah-Zadeh M, et al. BCL-3 expression promotes colorectal tumorigenesis through activation of AKT signalling. Gut. 2016;65:1151–64.

    CAS  Article  PubMed Central  Google Scholar 

  28. 28.

    Brasier AR, Lu M, Hai T, Lu Y, Boldogh I. NF-kappa B-inducible BCL-3 expression is an autoregulatory loop controlling nuclear p50/NF-kappa B1 residence. J Biol Chem. 2001;276:32080–93.

    CAS  Article  PubMed Central  Google Scholar 

  29. 29.

    Liu S, Sun X, Wang M, Hou Y, Zhan Y, Jiang Y, et al. A microRNA 221- and 222-mediated feedback loop maintains constitutive activation of NFkappaB and STAT3 in colorectal cancer cells. Gastroenterology. 2014;147:847–59 e11.

    CAS  Article  PubMed Central  Google Scholar 

  30. 30.

    Wang Y, Xu J, Gao G, Li J, Huang H, Jin H, et al. Tumor-suppressor NFkappaB2p100 interacts with ERK2 and stabilizes PTEN mRNA via inhibition of miR-494. Oncogene. 2016;35:4080–90.

    CAS  Article  Google Scholar 

  31. 31.

    Muller JR, Siebenlist U. Lymphotoxin beta receptor induces sequential activation of distinct NF-kappa B factors via separate signaling pathways. J Biol Chem. 2003;278:12006–12.

    Article  Google Scholar 

  32. 32.

    Wang S, Liu Z, Wang L, Zhang X. NF-kappaB signaling pathway, inflammation and colorectal cancer. Cell Mol Immunol. 2009;6:327–34.

    CAS  Article  PubMed Central  Google Scholar 

  33. 33.

    Wen D, Nong Y, Morgan JG, Gangurde P, Bielecki A, Dasilva J, et al. A selective small molecule IkappaB Kinase beta inhibitor blocks nuclear factor kappaB-mediated inflammatory responses in human fibroblast-like synoviocytes, chondrocytes, and mast cells. J Pharmacol Exp Ther. 2006;317:989–1001.

    CAS  Article  Google Scholar 

  34. 34.

    Vallabhapurapu S, Karin M. Regulation and function of NF-kappaB transcription factors in the immune system. Annu Rev Immunol. 2009;27:693–733.

    CAS  Article  Google Scholar 

  35. 35.

    Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144:646–74.

    CAS  Article  PubMed Central  Google Scholar 

  36. 36.

    Schumm K, Rocha S, Caamano J, Perkins ND. Regulation of p53 tumour suppressor target gene expression by the p52 NF-kappaB subunit. EMBO J. 2006;25:4820–32.

    CAS  Article  PubMed Central  Google Scholar 

  37. 37.

    Zhang J, Warren MA, Shoemaker SF, Ip MM. NFkappaB1/p50 is not required for tumor necrosis factor-stimulated growth of primary mammary epithelial cells: implications for NFkappaB2/p52 and RelB. Endocrinology. 2007;148:268–78.

    CAS  Article  Google Scholar 

  38. 38.

    Meyer N, Penn LZ. Reflecting on 25 years with MYC. Nat Rev Cancer. 2008;8:976–90.

    CAS  Article  Google Scholar 

  39. 39.

    Amati B, Alevizopoulos K, Vlach J. Myc and the cell cycle. Front Biosci. 1998;3:d250–68.

    CAS  Article  Google Scholar 

  40. 40.

    Chandramohan V, Mineva ND, Burke B, Jeay S, Wu M, Shen J, et al. c-Myc represses FOXO3a-mediated transcription of the gene encoding thep27(Kip1) cyclin dependent kinase inhibitor. J Cell Biochem. 2008;104:2091–106.

    CAS  Article  Google Scholar 

  41. 41.

    Perez-Roger I, Solomon DL, Sewing A, Land H. Myc activation of cyclin E/Cdk2 kinase involves induction of cyclin E gene transcription and inhibition ofp27(Kip1) binding to newly formed complexes. Oncogene. 1997;14:2373–81.

    CAS  Article  Google Scholar 

  42. 42.

    Bretones G, Acosta JC, Caraballo JM, Ferrandiz N, Gomez-Casares MT, Albajar M, et al. SKP2 oncogene is a direct MYC target gene and MYC down-regulatesp27(KIP1) through SKP2 in human leukemia cells. J Biol Chem. 2011;286:9815–25.

    CAS  Article  PubMed Central  Google Scholar 

  43. 43.

    Xue G, Yan HL, Zhang Y, Hao LQ, Zhu XT, Mei Q, et al. c-Myc-mediated repression of miR-15-16 in hypoxia is induced by increased HIF-2alpha and promotes tumor angiogenesis and metastasis by upregulating FGF2. Oncogene. 2015;34:1393–406.

    CAS  Article  Google Scholar 

  44. 44.

    Chen C, Cai S, Wang G, Cao X, Yang X, Luo X, et al. c-Myc enhances colon cancer cell-mediated angiogenesis through the regulation of HIF-1alpha. Biochem Biophys Res Commun. 2013;430:505–11.

    CAS  Article  Google Scholar 

  45. 45.

    Saamarthy K, Bjorner S, Johansson M, Landberg G, Massoumi R, Jirstrom K, et al. Early diagnostic value of Bcl-3 localization in colorectal cancer. BMC Cancer. 2015;15:341.

    Article  PubMed Central  Google Scholar 

  46. 46.

    Franzoso G, Carlson L, Poljak L, Shores EW, Epstein S, Leonardi A, et al. Mice deficient in nuclear factor (NF)-kappa B/p52 present with defects in humoral responses, germinal center reactions, and splenic microarchitecture. J Exp Med. 1998;187:147–59.

    CAS  Article  PubMed Central  Google Scholar 

  47. 47.

    Weih F, Caamano J. Regulation of secondary lymphoid organ development by the nuclear factor-kappaB signal transduction pathway. Immunol Rev. 2003;195:91–105.

    CAS  Article  Google Scholar 

  48. 48.

    Poljak L, Carlson L, Cunningham K, Kosco-Vilbois MH, Siebenlist U. Distinct activities of p52/NF-kappa B required for proper secondary lymphoid organ microarchitecture: functions enhanced by Bcl-3. J Immunol. 1999;163:6581–8.

    CAS  PubMed  Google Scholar 

  49. 49.

    Zhang X, Wang H, Claudio E, Brown K, Siebenlist U. A role for the IkappaB family member Bcl-3 in the control of central immunologic tolerance. Immunity. 2007;27:438–52.

    CAS  Article  PubMed Central  Google Scholar 

  50. 50.

    Franzoso G, Carlson L, Scharton-Kersten T, Shores EW, Epstein S, Grinberg A, et al. Critical roles for the Bcl-3 oncoprotein in T cell-mediated immunity, splenic microarchitecture, and germinal center reactions. Immunity. 1997;6:479–90.

    CAS  Article  Google Scholar 

  51. 51.

    Horton RM, Cai ZL, Ho SN, Pease LR. Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction. Biotechniques. 1990;8:528–35.

    CAS  PubMed  Google Scholar 

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This work was supported by the National Program on Key Research (2016YFC1302400), the National Basic Research Program (2014CB541904, 2014CB943600), National Natural Science Foundation of China (91742113, 31570902, 31370881), and Natural Science Foundation of Shanghai (14ZR1426300, 18ZR1424400, 18ZR1446400).

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Correspondence to Mingliang Wang or Xiaoren Zhang.

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These authors contributed equally: Yu Tao, Zhanjie Liu, Yingyong Hou.

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Tao, Y., Liu, Z., Hou, Y. et al. Alternative NF-κB signaling promotes colorectal tumorigenesis through transcriptionally upregulating Bcl-3. Oncogene 37, 5887–5900 (2018).

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