Skip to main content

Advertisement

SpringerLink
Log in

Clinicopathological significance of KU70/KU80, a key DNA damage repair protein in breast cancer

  • Preclinical study
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Although the role of BRCA1 and the homologous recombination (HR) pathway in breast cancer (BC) has been extensively studied, the alternative repair pathway for DNA double-strand breaks (DSBs), non-homologous end-joining (NHEJ) remains to be defined. Ku proteins bind to DNA DSB ends and play a key role in NHEJ. In this study we aimed to assess the expression and biological significance of the KU70/KU80 heterodimer in the different molecular classes of BC. The expression of KU70/KU80 was assessed immunohistochemically in a well-characterised and annotated series of 1302 unselected invasive BC cases with a long-term follow-up together with 25 cases with known BRCA1 mutations. The results were correlated with clinicopathological parameters, other DNA repair proteins and patient outcome. The expression of KU70/KU80 protein was further evaluated in various BC cell lines using western blotting and reverse-phase protein microarray (RPPA). Nuclear KU70/KU80 expression was correlated with features of poor prognosis including higher histological grade, lymphovascular invasion, negative oestrogen receptor expression, basal-like phenotype, P53 and CHK1 positivity. KU70/KU80 was expressed in all BRCA1-associated tumours and showed an inverse correlation with nuclear BRCA1 protein and aberrant cytoplasmic RAD51 expression. RPPA confirmed these results and showed higher expression of KU70/KU80 in BRCA1-deficient cell line compared to BRCA1-proficient cell line. KU70/KU80 expression showed an association with disease-free interval; however, it was not an independent predictor of outcome. As a conclusion, KU70/KU80 may play a role in DNA DSBs repair in HR-deficient tumours. Further study of other NHEJ markers in sporadic BC is warranted.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Khanna KK, Jackson SP (2001) DNA double-strand breaks: signaling, repair and the cancer connection. Nat Genet 27(3):247–254

    Article  PubMed  CAS  Google Scholar 

  2. O’Driscoll M, Jeggo PA (2006) The role of double-strand break repair—insights from human genetics. Nat Rev Genet 7(1):45–54

    Article  PubMed  Google Scholar 

  3. Scully R, Chen J, Plug A, Xiao Y, Weaver D, Feunteun J, Ashley T, Livingston DM (1997) Association of BRCA1 with Rad51 in mitotic and meiotic cells. Cell 88(2):265–275

    Article  PubMed  CAS  Google Scholar 

  4. Chen F, Wagner PD (1998) Pertussis toxin modification of PC12 cells lowers cytoskeletal F-actin and enhances norepinephrine secretion: involvement of protein kinase C and protein phosphatases. Arch Physiol Biochem 105(4):317–328

    PubMed  CAS  Google Scholar 

  5. Liu X, Holstege H, van der Gulden H, Treur-Mulder M, Zevenhoven J, Velds A, Kerkhoven RM, van Vliet MH, Wessels LF, Peterse JL et al (2007) Somatic loss of BRCA1 and p53 in mice induces mammary tumors with features of human BRCA1-mutated basal-like breast cancer. Proc Natl Acad Sci USA 104(29):12111–12116

    Article  PubMed  CAS  Google Scholar 

  6. Foulkes WD, Stefansson IM, Chappuis PO, Begin LR, Goffin JR, Wong N, Trudel M, Akslen LA (2003) Germline BRCA1 mutations and a basal epithelial phenotype in breast cancer. J Natl Cancer Inst 95(19):1482–1485

    Article  PubMed  CAS  Google Scholar 

  7. Turner NC, Reis-Filho JS, Russell AM, Springall RJ, Ryder K, Steele D, Savage K, Gillett CE, Schmitt FC, Ashworth A et al (2007) BRCA1 dysfunction in sporadic basal-like breast cancer. Oncogene 26(14):2126–2132

    Article  PubMed  CAS  Google Scholar 

  8. Rakha EA, El-Sheikh SE, Kandil MA, El-Sayed ME, Green AR, Ellis IO (2008) Expression of BRCA1 protein in breast cancer and its prognostic significance. Hum Pathol 39(6):857–865

    Article  PubMed  CAS  Google Scholar 

  9. Rakha EA, Reis-Filho JS, Ellis IO (2008) Basal-like breast cancer: a critical review. J Clin Oncol 26(15):2568–2581

    Article  PubMed  Google Scholar 

  10. McCabe N, Turner NC, Lord CJ, Kluzek K, Bialkowska A, Swift S, Giavara S, O’Connor MJ, Tutt AN, Zdzienicka MZ et al (2006) Deficiency in the repair of DNA damage by homologous recombination and sensitivity to poly(ADP-ribose) polymerase inhibition. Cancer Res 66(16):8109–8115

    Article  PubMed  CAS  Google Scholar 

  11. Burma S, Chen BP, Chen DJ (2006) Role of non-homologous end joining (NHEJ) in maintaining genomic integrity. DNA Repair (Amst) 5(9–10):1042–1048

    Article  CAS  Google Scholar 

  12. Lieber MR (2008) The mechanism of human nonhomologous DNA end joining. J Biol Chem 283(1):1–5

    Article  PubMed  CAS  Google Scholar 

  13. Lieber MR (2010) The mechanism of double-strand DNA break repair by the nonhomologous DNA end-joining pathway. Annu Rev Biochem 79:181–211

    Article  PubMed  CAS  Google Scholar 

  14. Downs JA, Jackson SP (2004) A means to a DNA end: the many roles of Ku. Nat Rev Mol Cell Biol 5(5):367–378

    Article  PubMed  CAS  Google Scholar 

  15. Chen Y, Liu Z, Zhu K, Wu Y, Yin H (2008) Investigation of the relationship between DNA-dependent protein kinase and lymphatic metastasis in colorectal cancer. Chin-Ger J Clin Oncol 7(6):348–351

    Article  Google Scholar 

  16. Komuro Y, Watanabe T, Hosoi Y, Matsumoto Y, Nakagawa K, Tsuno N, Kazama S, Kitayama J, Suzuki N, Nagawa H (2002) The expression pattern of Ku correlates with tumor radiosensitivity and disease free survival in patients with rectal carcinoma. Cancer 95(6):1199–1205

    Article  PubMed  Google Scholar 

  17. Madjd Z, Pinder SE, Paish C, Ellis IO, Carmichael J, Durrant LG (2003) Loss of CD59 expression in breast tumours correlates with poor survival. J Pathol 200(5):633–639

    Article  PubMed  CAS  Google Scholar 

  18. Rakha EA, Elsheikh SE, Aleskandarany MA, Habashi HO, Green AR, Powe DG, El-Sayed ME, Benhasouna A, Brunet JS, Akslen LA et al (2009) Triple-negative breast cancer: distinguishing between basal and nonbasal subtypes. Clin Cancer Res 15(7):2302–2310

    Article  PubMed  CAS  Google Scholar 

  19. Camp RL, Dolled-Filhart M, Rimm DL (2004) X-tile a new bio-informatics tool for biomarker assessment and outcome-based cut-point optimization. Clin Cancer Res 10(21):7252–7259

    Article  PubMed  CAS  Google Scholar 

  20. Moynahan ME, Cui TY, Jasin M (2001) Homology-directed dna repair, mitomycin-c resistance, and chromosome stability is restored with correction of a Brca1 mutation. Cancer Res 61(12):4842–4850

    PubMed  CAS  Google Scholar 

  21. Mimori T, Hardin JA (1986) Mechanism of interaction between Ku protein and DNA. J Biol Chem 261(22):10375–10379

    PubMed  CAS  Google Scholar 

  22. Rathmell WK, Chu G (1994) Involvement of the Ku autoantigen in the cellular response to DNA double-strand breaks. Proc Natl Acad Sci USA 91(16):7623–7627

    Article  PubMed  CAS  Google Scholar 

  23. Finnie NJ, Gottlieb TM, Blunt T, Jeggo PA, Jackson SP (1995) DNA-dependent protein kinase activity is absent in xrs-6 cells: implications for site-specific recombination and DNA double-strand break repair. Proc Natl Acad Sci USA 92(1):320–324

    Article  PubMed  CAS  Google Scholar 

  24. Lees-Miller SP, Godbout R, Chan DW, Weinfeld M, Day RS 3rd, Barron GM, Allalunis-Turner J (1995) Absence of p350 subunit of DNA-activated protein kinase from a radiosensitive human cell line. Science 267(5201):1183–1185

    Article  PubMed  CAS  Google Scholar 

  25. Adachi N, Ishino T, Ishii Y, Takeda S, Koyama H (2001) DNA ligase IV-deficient cells are more resistant to ionizing radiation in the absence of Ku70: implications for DNA double-strand break repair. Proc Natl Acad Sci 98(21):12109

    Article  PubMed  CAS  Google Scholar 

  26. Bau DT, Fu YP, Chen ST, Cheng TC, Yu JC, Wu PE, Shen CY (2004) Breast cancer risk and the DNA double-strand break end-joining capacity of nonhomologous end-joining genes are affected by BRCA1. Cancer Res 64(14):5013

    Article  PubMed  CAS  Google Scholar 

  27. Ghezzi TL, Brum IS, Biolchi V, Garicochea B, Fillmann LS, Corleta OC (2011) Is there any association between TACSTD2, KIAA1253, Ku70 and mutant KRAS gene expression and clinical-pathological features of colorectal cancer? Exp Oncol 33(1):28–32

    PubMed  CAS  Google Scholar 

  28. Martin RW, Orelli BJ, Yamazoe M, Minn AJ, Takeda S, Bishop DK (2007) RAD51 up-regulation bypasses BRCA1 function and is a common feature of BRCA1-deficient breast tumors. Cancer Res 67(20):9658–9665

    Article  PubMed  CAS  Google Scholar 

  29. Soderlund Leifler K, Queseth S, Fornander T, Askmalm MS (2010) Low expression of Ku70/80, but high expression of DNA-PKcs, predict good response to radiotherapy in early breast cancer. Int J Oncol 37(6):1547–1554

    PubMed  Google Scholar 

  30. Pavon MA, Parreno M, Leon X, Sancho FJ, Cespedes MV, Casanova I, Lopez-Pousa A, Mangues MA, Quer M, Barnadas A et al (2008) Ku70 predicts response and primary tumor recurrence after therapy in locally advanced head and neck cancer. Int J Cancer 123(5):1068–1079

    Article  PubMed  CAS  Google Scholar 

  31. Grabsch H, Dattani M, Barker L, Maughan N, Maude K, Hansen O, Gabbert HE, Quirke P, Mueller W (2006) Expression of DNA double-strand break repair proteins ATM and BRCA1 predicts survival in colorectal cancer. Clin Cancer Res 12(5):1494–1500

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

Conflict of interest

The authors declared that they have no conflict of interest

Author information

Authors and Affiliations

  1. Department of Histopathology and School of Molecular Medical Sciences, The University of Nottingham and Nottingham University Hospitals NHS Trust, Nottingham City Hospital, Nottingham, UK

    Alaa T. Alshareeda, Andrew R. Green, Christopher Nolan, Ahmed benHasouna, Ian O. Ellis & Emad A. Rakha

  2. Ministry of Higher Education, Riyadh, Kingdom of Saudi Arabia

    Alaa T. Alshareeda

  3. Immunology, School of Molecular Medical Sciences, University of Nottingham, Nottingham, UK

    Ola H. Negm & Paddy Tighe

  4. Medical Microbiology and Immunology Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt

    Ola H. Negm

  5. Department of Oncology and School of Molecular Medical Sciences, The University of Nottingham and Nottingham University Hospitals NHS Trust, Nottingham City Hospital, Nottingham, UK

    Nada Albarakati, Rebeka Sultana & Srinivasan Madhusudan

  6. Breast Cancer Pathology Research Group, School of Molecular Medical Sciences, Academic Unit of Clinical Oncology, University of Nottingham, Nottingham City Hospital, Hucknall Road, Nottingham, NG5 1PB, UK

    Alaa T. Alshareeda

Authors
  1. Alaa T. Alshareeda
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ola H. Negm
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nada Albarakati
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Andrew R. Green
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Christopher Nolan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rebeka Sultana
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Srinivasan Madhusudan
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ahmed benHasouna
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Paddy Tighe
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ian O. Ellis
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Emad A. Rakha
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alaa T. Alshareeda.

Additional information

Ola H. Negm is first joint author.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Alshareeda, A.T., Negm, O.H., Albarakati, N. et al. Clinicopathological significance of KU70/KU80, a key DNA damage repair protein in breast cancer. Breast Cancer Res Treat 139, 301–310 (2013). https://doi.org/10.1007/s10549-013-2542-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-013-2542-x

Keywords

Search

Navigation