Breast Cancer Research and Treatment

, Volume 127, Issue 1, pp 53–63 | Cite as

DEAD box 1: a novel and independent prognostic marker for early recurrence in breast cancer

  • Devon R. Germain
  • Kathryn Graham
  • Darryl D. Glubrecht
  • Judith C. Hugh
  • John R. Mackey
  • Roseline Godbout
Preclinical study


Breast cancer is a heterogeneous disease characterized by diverse molecular signatures and a variable response to therapy. Clinical management of breast cancer is guided by the expression of estrogen and progesterone receptors and HER2 amplification. New prognostic and predictive markers, as well as additional targets for therapy, are needed for more effective management of this disease. Gene expression microarrays were probed with RNAs from 176 primary breast cancer samples and tissue microarrays immunostained with anti-DDX1 antibody, an antibody to DEAD box protein DDX1, a putative RNA-RNA and RNA-DNA unwinding protein normally found in the nucleus. Half of the patient cohort had experienced early relapse despite standard adjuvant therapy, but were otherwise matched for estrogen receptor and HER2 status, stage and duration of follow-up. Here, we identify DDX1 RNA overexpression as an independent prognostic marker for early recurrence in primary breast cancer, with a hazard ratio of 4.31 based on logrank analysis of Kaplan–Meier curves. Elevated levels of DDX1 protein in the cytoplasm also independently correlate with early recurrence with a hazard ratio of 1.90. In conclusion, our data indicate a strong and independent association between poor prognosis and deregulation of the DEAD box protein DDX1. We propose that elevated levels of DDX1 RNA or the presence of DDX1 in the cytoplasm could serve as an effective prognostic biomarker for early recurrence in primary breast cancer.


Breast cancer DEAD box 1 Gene expression Tissue microarray Prognostic marker Early recurrence 



We are grateful to Lillian Cook and Cheryl Santos for their excellent technical assistance, Adrian Driga for his help with database analysis and Sunita Ghosh for her assistance with statistical analyses. This study was supported by grants from the Alberta Cancer Board—Research Initiative Program (R.G.), Alberta Cancer Research Institute (J.R.M.) and an Alberta Cancer Foundation studentship (D.R.G.).


  1. 1.
    Porter P (2008) “Westernizing” women’s risks? Breast cancer in lower-income countries. N Engl J Med 358:213–216PubMedCrossRefGoogle Scholar
  2. 2.
    Jones SE (2008) Metastatic breast cancer: the treatment challenge. Clin Breast Cancer 8:224–233PubMedCrossRefGoogle Scholar
  3. 3.
    Bergh J, Jonsson PE, Glimelius B, Nygren P (2001) A systematic overview of chemotherapy effects in breast cancer. Acta Oncol 40:253–281PubMedCrossRefGoogle Scholar
  4. 4.
    Lopez-Tarruella S, Martin M (2009) Recent advances in systemic therapy: advances in adjuvant systemic chemotherapy of early breast cancer. Breast Cancer Res 11:204PubMedCrossRefGoogle Scholar
  5. 5.
    Barnadas A, Gil M, Sanchez-Rovira P, Llombart A, Adrover E, Estevez LG, De la Haba J, Calvo L (2008) Neoadjuvant endocrine therapy for breast cancer: past, present and future. Anticancer Drugs 19:339–347PubMedCrossRefGoogle Scholar
  6. 6.
    Fossati R, Confalonieri C, Torri V, Ghislandi E, Penna A, Pistotti V, Tinazzi A, Liberati A (1998) Cytotoxic and hormonal treatment for metastatic breast cancer: a systematic review of published randomized trials involving 31,510 women. J Clin Oncol 16:3439–3460PubMedGoogle Scholar
  7. 7.
    Cheang MC, Chia SK, Voduc D, Gao D, Leung S, Snider J, Watson M, Davies S, Bernard PS, Parker JS, Perou CM, Ellis MJ, Nielsen TO (2009) Ki67 index, HER2 status, and prognosis of patients with luminal B breast cancer. J Natl Cancer Inst 101:736–750PubMedCrossRefGoogle Scholar
  8. 8.
    Fan C, Oh DS, Wessels L, Weigelt B, Nuyten DS, Nobel AB, van’t Veer LJ, Perou CM (2006) Concordance among gene-expression-based predictors for breast cancer. N Engl J Med 355:560–569PubMedCrossRefGoogle Scholar
  9. 9.
    Perou CM, Sorlie T, Eisen MB, van de Rijn M, Jeffrey SS, Rees CA, Pollack JR, Ross DT, Johnsen H, Akslen LA, Fluge O, Pergamenschikov A, Williams C, Zhu SX, Lonning PE, Borresen-Dale AL, Brown PO, Botstein D (2000) Molecular portraits of human breast tumours. Nature 406:747–752PubMedCrossRefGoogle Scholar
  10. 10.
    Hugh J, Hanson J, Cheang MC, Nielsen TO, Perou CM, Dumontet C, Reed J, Krajewska M, Treilleux I, Rupin M, Magherini E, Mackey J, Martin M, Vogel C (2009) Breast cancer subtypes and response to docetaxel in node-positive breast cancer: use of an immunohistochemical definition in the BCIRG 001 trial. J Clin Oncol 27:1168–1176PubMedCrossRefGoogle Scholar
  11. 11.
    Kreike B, van Kouwenhove M, Horlings H, Weigelt B, Peterse H, Bartelink H, van de Vijver MJ (2007) Gene expression profiling and histopathological characterization of triple-negative/basal-like breast carcinomas. Breast Cancer Res 9:R65PubMedCrossRefGoogle Scholar
  12. 12.
    Brenton JD, Carey LA, Ahmed AA, Caldas C (2005) Molecular classification and molecular forecasting of breast cancer: ready for clinical application? J Clin Oncol 23:7350–7360PubMedCrossRefGoogle Scholar
  13. 13.
    Chiuri VE, Leo G, Lorusso V (2007) Clinical and therapeutic perspectives of gene expression profiling for breast cancer. Ann Oncol 18(Suppl 6):Vi58–Vi62PubMedCrossRefGoogle Scholar
  14. 14.
    Oakman C, Bessi S, Zafarana E, Galardi F, Biganzoli L, Di Leo A (2009) Recent advances in systemic therapy: new diagnostics and biological predictors of outcome in early breast cancer. Breast Cancer Res 11:205PubMedCrossRefGoogle Scholar
  15. 15.
    Stadler ZK, Come SE (2009) Review of gene-expression profiling and its clinical use in breast cancer. Crit Rev Oncol Hematol 69:1–11PubMedCrossRefGoogle Scholar
  16. 16.
    Slodkowska EA, Ross JS (2009) MammaPrint 70-gene signature: another milestone in personalized medical care for breast cancer patients. Expert Rev Mol Diagn 9:417–422PubMedCrossRefGoogle Scholar
  17. 17.
    Albain KS, Barlow WE, Shak S, Hortobagyi GN, Livingston RB, Yeh IT, Ravdin P, Bugarini R, Baehner FL, Davidson NE, Sledge GW, Winer EP, Hudis C, Ingle JN, Perez EA, Pritchard KI, Shepherd L, Gralow JR, Yoshizawa C, Allred DC, Osborne CK, Hayes DF (2010) Prognostic and predictive value of the 21-gene recurrence score assay in postmenopausal women with node-positive, oestrogen-receptor-positive breast cancer on chemotherapy: a retrospective analysis of a randomised trial. Lancet Oncol 11:55–65PubMedCrossRefGoogle Scholar
  18. 18.
    Mook S, Schmidt MK, Weigelt B, Kreike B, Eekhout I, van de Vijver MJ, Glas AM, Floore A, Rutgers EJ, van ‘t Veer LJ (2009) The 70-gene prognosis signature predicts early metastasis in breast cancer patients between 55 and 70 years of age. Ann Oncol. doi: 10.1093/annonc/mdp388
  19. 19.
    Linder P (2006) Dead-box proteins: a family affair—active and passive players in RNP-remodeling. Nucleic Acids Res 34:4168–4180PubMedCrossRefGoogle Scholar
  20. 20.
    Godbout R, Squire J (1993) Amplification of a DEAD box protein gene in retinoblastoma cell lines. Proc Natl Acad Sci USA 90:7578–7582PubMedCrossRefGoogle Scholar
  21. 21.
    Squire JA, Thorner PS, Weitzman S, Maggi JD, Dirks P, Doyle J, Hale M, Godbout R (1995) Co-amplification of MYCN and a DEAD box gene (DDX1) in primary neuroblastoma. Oncogene 10:1417–1422PubMedGoogle Scholar
  22. 22.
    Manohar CF, Salwen HR, Brodeur GM, Cohn SL (1995) Co-amplification and concomitant high levels of expression of a DEAD box gene with MYCN in human neuroblastoma. Genes Chromosomes Cancer 14:196–203PubMedCrossRefGoogle Scholar
  23. 23.
    George RE, Kenyon RM, McGuckin AG, Malcolm AJ, Pearson AD, Lunec J (1996) Investigation of co-amplification of the candidate genes ornithine decarboxylase, ribonucleotide reductase, syndecan-1 and a DEAD box gene, DDX1, with N-myc in neuroblastoma. United Kingdom Children’s Cancer Study Group. Oncogene 12:1583–1587PubMedGoogle Scholar
  24. 24.
    Tanaka K, Okamoto S, Ishikawa Y, Tamura H, Hara T (2009) DDX1 is required for testicular tumorigenesis, partially through the transcriptional activation of 12p stem cell genes. Oncogene 28:2142–2151PubMedCrossRefGoogle Scholar
  25. 25.
    Zinsmaier KE, Eberle KK, Buchner E, Walter N, Benzer S (1994) Paralysis and early death in cysteine string protein mutants of Drosophila. Science 263:977–980PubMedCrossRefGoogle Scholar
  26. 26.
    Bleoo S, Sun X, Hendzel MJ, Rowe JM, Packer M, Godbout R (2001) Association of human DEAD box protein DDX1 with a cleavage stimulation factor involved in 3′-end processing of pre-MRNA. Mol Biol Cell 12:3046–3059PubMedGoogle Scholar
  27. 27.
    Kanai Y, Dohmae N, Hirokawa N (2004) Kinesin transports RNA: isolation and characterization of an RNA-transporting granule. Neuron 43:513–525PubMedCrossRefGoogle Scholar
  28. 28.
    Fang J, Kubota S, Yang B, Zhou N, Zhang H, Godbout R, Pomerantz RJ (2004) A DEAD box protein facilitates HIV-1 replication as a cellular co-factor of Rev. Virology 330:471–480PubMedCrossRefGoogle Scholar
  29. 29.
    Li L, Monckton EA, Godbout R (2008) A role for DEAD box 1 at DNA double-strand breaks. Mol Cell Biol 28:6413–6425PubMedCrossRefGoogle Scholar
  30. 30.
    Li L, Roy K, Katyal S, Sun X, Bleoo S, Godbout R (2006) Dynamic nature of cleavage bodies and their spatial relationship to DDX1 bodies, Cajal bodies, and gems. Mol Biol Cell 17:1126–1140PubMedCrossRefGoogle Scholar
  31. 31.
    Godbout R, Packer M, Bie W (1998) Overexpression of a DEAD box protein (DDX1) in neuroblastoma and retinoblastoma cell lines. J Biol Chem 273:21161–21168PubMedCrossRefGoogle Scholar
  32. 32.
    McShane LM, Altman DG, Sauerbrei W, Taube SE, Gion M, Clark GM (2006) REporting recommendations for tumor MARKer prognostic studies (REMARK). Breast Cancer Res Treat 100:229–235PubMedCrossRefGoogle Scholar
  33. 33.
    Early Breast Cancer Trialists’ Collaborative Group (EBCTCG) (2005) Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. Lancet 365:1687–1717Google Scholar
  34. 34.
    Sunden Y, Semba S, Suzuki T, Okada Y, Orba Y, Nagashima K, Umemura T, Sawa H (2007) Identification of DDX1 as a JC virus transcriptional control region-binding protein. Microbiol Immunol 51:327–337PubMedCrossRefGoogle Scholar
  35. 35.
    Bordeleau ME, Matthews J, Wojnar JM, Lindqvist L, Novac O, Jankowsky E, Sonenberg N, Northcote P, Teesdale-Spittle P, Pelletier J (2005) Stimulation of mammalian translation initiation factor eIF4A activity by a small molecule inhibitor of eukaryotic translation. Proc Natl Acad Sci USA 102:10460–10465PubMedCrossRefGoogle Scholar
  36. 36.
    Erkizan HV, Kong Y, Merchant M, Schlottmann S, Barber-Rotenberg JS, Yuan L, Abaan OD, Chou TH, Dakshanamurthy S, Brown ML, Uren A, Toretsky JA (2009) A small molecule blocking oncogenic protein EWS-FLI1 interaction with RNA helicase A inhibits growth of Ewing’s sarcoma. Nat Med 15:750–756PubMedCrossRefGoogle Scholar
  37. 37.
    Fuller-Pace FV, Ali S (2008) The DEAD box RNA helicases p68 (Ddx5) and p72 (Ddx17): novel transcriptional co-regulators. Biochem Soc Trans 36:609–612PubMedCrossRefGoogle Scholar
  38. 38.
    Wortham NC, Ahamed E, Nicol SM, Thomas RS, Periyasamy M, Jiang J, Ochocka AM, Shousha S, Huson L, Bray SE, Coombes RC, Ali S, Fuller-Pace FV (2009) The DEAD-box protein p72 regulates ERalpha-/oestrogen-dependent transcription and cell growth, and is associated with improved survival in ERalpha-positive breast cancer. Oncogene. doi:  10.1038/onc.2009.261
  39. 39.
    Lin F, Wang R, Shen JJ, Wang X, Gao P, Dong K, Zhang HZ (2008) Knockdown of RCK/p54 expression by RNAi inhibits proliferation of human colorectal cancer cells in vitro and in vivo. Cancer Biol Ther 7:1669–1676PubMedGoogle Scholar
  40. 40.
    Cho B, Lim Y, Lee DY, Park SY, Lee H, Kim WH, Yang H, Bang YJ, Jeoung DI (2002) Identification and characterization of a novel cancer/testis antigen gene CAGE. Biochem Biophys Res Commun 292:715–726PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Devon R. Germain
    • 1
  • Kathryn Graham
    • 1
  • Darryl D. Glubrecht
    • 1
  • Judith C. Hugh
    • 2
  • John R. Mackey
    • 1
  • Roseline Godbout
    • 1
  1. 1.Department of Oncology, School of Cancer/Engineering/Imaging SciencesUniversity of Alberta, Cross Cancer InstituteEdmontonCanada
  2. 2.Department of Lab Medicine and PathologyUniversity of AlbertaEdmontonCanada

Personalised recommendations