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PARP1 and phospho-p65 protein expression is increased in human HER2-positive breast cancers

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Abstract

Previous studies have shown that basal breast cancers, which may have an inherent “BRCAness” phenotype and sensitivity to inhibitors of poly (ADP-Ribose) polymerase (PARP), express elevated levels of PARP1. Our lab recently reported that HER2+ breast cancers also exhibit sensitivity to PARP inhibitors (PARPi) by attenuating the NF-κB pathway. In this study, we assessed PARP1 and phospho-p65, a marker of activated NF-κB levels in human breast cancer tissues. PARP1 and PARP2 copy number, mRNA, and protein expression was assessed by interrogating the PAM-50 defined breast cancer patient set from the TCGA using cBioPortal. PARP1 and phospho-p65 immunohistochemistry and correlation to clinical parameters was conducted using 307 primary breast cancer specimens (132 basal, 82 luminal, 93 HER2+) through univariate and multivariate analyses. In the PAM50 breast cancer data set, PARP1 and 2 expression was altered in 24/58 (41 %) HER2+, 32/81 (40 %) basal, and 75/324 (23 %) luminal A/B breast cancer patients. This correlated with a statistically significant increase in PARP1 protein levels in HER2+ and basal but not luminal breast cancers (p = 0.003, p = 0.027, p = 0.289, respectively). No change in PARP2 protein level was observed. Interestingly, using breast cancer specimens from 307 patients, HER2 positivity correlated with elevated PARP1 expression (p < 0.0001) and was three times more likely than HER2 negative breast cancers to exhibit high PARP1 levels. No significant differences were noted between race, ER status, or PR status for PARP1 expression. Additionally, we found a significant correlation between HER2 status and phospho-p65 expression (p < 0.0001). Lastly, a direct correlation between PARP1 and phospho-p65 (p < 0.0001) was noted. These results indicate a potential connection between HER2, PARP1, and phospho-p65. Furthermore, these data suggest that the PARPi sensitivity we previously observed in HER2+ breast cancer cells may be due to elevated PARP1 expression.

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References

  1. Perou CM et al (2000) Molecular portraits of human breast tumours. Nature 406(6797):747–752

    Article  CAS  PubMed  Google Scholar 

  2. Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144(5):646–674

    Article  CAS  PubMed  Google Scholar 

  3. Bryant HE et al (2005) Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase. Nature 434(7035):913–917

    Article  CAS  PubMed  Google Scholar 

  4. Farmer H et al (2005) Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature 434(7035):917–921

    Article  CAS  PubMed  Google Scholar 

  5. Fong PC et al (2009) Inhibition of poly(ADP-Ribose) polymerase in tumors from BRCA mutation carriers. N Engl J Med 361(2):123–134

    Article  CAS  PubMed  Google Scholar 

  6. Audeh MW et al (2010) Oral poly(ADP-ribose) polymerase inhibitor olaparib in patients with BRCA1 or BRCA2 mutations and recurrent ovarian cancer: a proof-of-concept trial. Lancet 376(9737):245–251

    Article  CAS  PubMed  Google Scholar 

  7. Tutt A et al (2010) Oral poly (ADP-ribose) polymerase inhibitor olaparib in patients with BRCA1 or BRCA2 mutations and advanced breast cancer: a proof-of-concept trial. Lancet 376(9737):235–244

    Article  CAS  PubMed  Google Scholar 

  8. Turner NC, Reis-Filho JS (2006) Basal-like breast cancer and the BRCA1 phenotype. Oncogene 25(43):5846–5853

    Article  CAS  PubMed  Google Scholar 

  9. Domagala P et al (2011) PARP-1 expression in breast cancer including BRCA1-associated, triple negative and basal-like tumors: possible implications for PARP-1 inhibitor therapy. Breast Cancer Res Treat 127(3):861–869

    Article  CAS  PubMed  Google Scholar 

  10. Nowsheen S et al (2012) HER2 overexpression renders human breast cancers sensitive to PARP inhibition independently of any defect in homologous recombination DNA repair. Cancer Res 72(18):4796–4806

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Hassa PO, Hottiger MO (1999) A role of poly (ADP-ribose) polymerase in NF-kappaB transcriptional activation. Biol Chem 380(7–8):953–959

    CAS  PubMed  Google Scholar 

  12. Hassa P, Hottiger M (2002) The functional role of poly (ADP-ribose) polymerase 1 as novel coactivator of NF-κB in inflammatory disorders. Cell Mol Life Sci CMLS 59(9):1534–1553

    Article  CAS  Google Scholar 

  13. O’Shaughnessy J et al (2011) Iniparib plus chemotherapy in metastatic triple-negative breast cancer. N Engl J Med 364(3):205–214

    Article  PubMed  Google Scholar 

  14. Rottenberg S et al (2008) High sensitivity of BRCA1-deficient mammary tumors to the PARP inhibitor AZD2281 alone and in combination with platinum drugs. Proc Natl Acad Sci 105(44):17079–17084

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Cancer Genome Atlas Network (2012) Comprehensive molecular portraits of human breast tumours. Nature 490(7418):61–70

    Article  Google Scholar 

  16. Biswas DK et al (2004) NF-kappa B activation in human breast cancer specimens and its role in cell proliferation and apoptosis. Proc Natl Acad Sci USA 101(27):10137–10142

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Biswas DK, Iglehart JD (2006) Linkage between EGFR family receptors and nuclear factor kappaB (NF-kappaB) signaling in breast cancer. J Cell Physiol 209(3):645–652

    Article  CAS  PubMed  Google Scholar 

  18. Izzo JG et al (2006) Pretherapy nuclear factor-kappaB status, chemoradiation resistance, and metastatic progression in esophageal carcinoma. Mol Cancer Ther 5(11):2844–2850

    Article  CAS  PubMed  Google Scholar 

  19. Braunstein S, Formenti SC, Schneider RJ (2008) Acquisition of stable inducible up-regulation of nuclear factor-kappaB by tumor necrosis factor exposure confers increased radiation resistance without increased transformation in breast cancer cells. Mol Cancer Res 6(1):78–88

    Article  CAS  PubMed  Google Scholar 

  20. Prasad S, Ravindran J, Aggarwal BB (2010) NF-kappaB and cancer: how intimate is this relationship. Mol Cell Biochem 336(1–2):25–37

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Merkhofer EC, Cogswell P, Baldwin AS (2010) Her2 activates NF-kappaB and induces invasion through the canonical pathway involving IKKalpha. Oncogene 29(8):1238–1248

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Singh S et al (2007) Nuclear factor-kappaB activation: a molecular therapeutic target for estrogen receptor-negative and epidermal growth factor receptor family receptor-positive human breast cancer. Mol Cancer Ther 6(7):1973–1982

    Article  CAS  PubMed  Google Scholar 

  23. Shapochka D et al (2012) Relationship between NF-κB, ER, PR, Her2/neu, Ki67, p53 expression in human breast cancer. Exp Oncol 34(4):358–363

    CAS  PubMed  Google Scholar 

  24. von Minckwitz G et al (2011) Cytoplasmic poly (adenosine diphosphate–ribose) polymerase expression is predictive and prognostic in patients with breast cancer treated with neoadjuvant chemotherapy. J Clin Oncol 29(16):2150–2157

    Article  Google Scholar 

  25. Ossovskaya V et al (2011) Exploring molecular pathways of triple-negative breast cancer. Genes Cancer 2(9):870–879

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Cao Y, Luo J-L, Karin M (2007) IκB kinase α kinase activity is required for self-renewal of ErbB2/Her2-transformed mammary tumor-initiating cells. Proc Natl Acad Sci 104(40):15852–15857

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Stilmann M et al (2009) A nuclear poly (ADP-ribose)-dependent signalosome confers DNA damage-induced IκB kinase activation. Mol Cell 36(3):365–378

    Article  CAS  PubMed  Google Scholar 

  28. Veuger SJ, Hunter JE, Durkacz BW (2009) Ionizing radiation-induced NF-kappaB activation requires PARP-1 function to confer radioresistance. Oncogene 28(6):832–842

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Dolcet X et al (2005) NF-κB in development and progression of human cancer. Virchows Arch 446(5):475–482

    Article  CAS  PubMed  Google Scholar 

  30. Guo RX et al (2008) Increased staining for phosphorylated AKT and nuclear factor-κB p65 and their relationship with prognosis in epithelial ovarian cancer. Pathol Int 58(12):749–756

    Article  CAS  PubMed  Google Scholar 

  31. Weichert W et al (2007) High expression of RelA/p65 is associated with activation of nuclear factor-κB-dependent signaling in pancreatic cancer and marks a patient population with poor prognosis. Br J Cancer 97(4):523–530

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  32. Sasaki N et al (2001) Nuclear factor-κB p65 (RelA) transcription factor is constitutively activated in human gastric carcinoma tissue. Clin Cancer Res 7(12):4136–4142

    CAS  PubMed  Google Scholar 

  33. Li W et al (2010) Constitutive activation of nuclear factor-kappa B (NF-κB) signaling pathway in fibrolamellar hepatocellular carcinoma. Int J Clin Exp Pathol 3(3):238

    PubMed Central  CAS  Google Scholar 

  34. Prusty BK, Husain SA, Das BC (2005) Constitutive activation of nuclear factor-κB: preferential homodimerization of p50 subunits in cervical carcinoma. Front Biosci 10(5):1510–1519

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work is supported in part by the Minority Biospecimen/Biobanking Geographic Management Program for Region 3 (BMaP-3) (3U54 CA153509-03S1) (ESY), AACR-Genentech BioOncology Career Development Award for Cancer Research on the HER Family Pathway (12-30-18-YANG, to ESY), Susan G. Komen Career Catalyst Award (CCR12364491, to ESY), NCI/H. Lee Moffitt Cancer Center BMaP Pilot Grant (10-16308-03-17-G1, to ESY), the UAB MSTP (NIH-NIGMS 5T32GM008361-21) (JAS), the UAB Cell and Molecular Biology Training grant (5T32GM008111-27) (JAS), and an intramural pilot grant from the Department of Radiation Oncology (to LK). This work was also supported by the Breast SPORE (5P50CA 089019) and Tissue Procurement Shared Facility (P30CA-13148-41) of the Comprehensive Cancer Center at UAB.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

The experiments in this manuscript comply with the current laws of the country in which they were performed.

Research involving human participants and/or animals

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This article does not contain any studies with animals performed by any of the authors.

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    Authors and Affiliations

    1. Department of Radiation Oncology, University of Alabama at Birmingham, 1700 6th Avenue South, 176F HSROC Rm 2232 N, Birmingham, AL, USA

      Jennifer Stanley, Lisa Klepczyk, Kimberly Keene, Monica Wielgos & Eddy S. Yang

    2. Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA

      Shi Wei, William Grizzle & Jason Brazelton

    3. Department of Preventitive Medicine, University of Alabama at Birmingham, Birmingham, AL, USA

      Yufeng Li

    4. Department of Hematology and Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

      Andres Forero

    5. Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA

      Albert F. LoBuglio

    6. Department of Cell, Developmental, and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, USA

      Eddy S. Yang

    7. Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, AL, USA

      Eddy S. Yang

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    1. Jennifer Stanley
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    2. Lisa Klepczyk
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    3. Kimberly Keene
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    Correspondence to Eddy S. Yang.

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    Jennifer Stanley and Lisa Klepczyk have contributed equally to this work.

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    Stanley, J., Klepczyk, L., Keene, K. et al. PARP1 and phospho-p65 protein expression is increased in human HER2-positive breast cancers. Breast Cancer Res Treat 150, 569–579 (2015). https://doi.org/10.1007/s10549-015-3359-6

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    • DOI: https://doi.org/10.1007/s10549-015-3359-6

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