Tumor Biology

, Volume 36, Issue 7, pp 5415–5423 | Cite as

USP2 promotes cell migration and invasion in triple negative breast cancer cell lines

  • Qing Qu
  • Yan Mao
  • Gang Xiao
  • Xiaochun Fei
  • Jinglong Wang
  • Yuzi Zhang
  • Junjun Liu
  • Guangcun Cheng
  • Xiaosong Chen
  • Jianhua Wang
  • Kunwei Shen
Research Article


Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer that is often associated with a poor prognosis. The aim of our study was to identify biomarkers predictive of TNBC progression. Primary TNBC breast tissue samples including four with metastasis and six without metastasis were subjected to Affymetrix GeneChip® analysis (human genome U133). Ubiquitin-specific protease 2 (USP2) was identified as an upregulated gene in the metastatic group, and its expression was analyzed by immunohistochemistry in 121 primary breast cancers, 13 paired normal tissues, and 13 paired metastatic lesions. Survival analysis was performed using the log-rank test and Cox regression hazard model. Matrigel migration and invasion assays in USP2-silenced and USP2-overexpressed breast cancer cell lines were used to investigate the mechanisms of USP2 in vitro. Positive immunostaining for USP2 was detected in breast tumors and was correlated with estrogen receptor (ER) and progesterone receptor (PR) statuses and TNBC subtype. USP2 was overexpressed in distant metastatic lesions compared with primary breast cancers. Survival analyses demonstrated that positive USP2 is a poor prognostic factor for disease-free survival. Silencing of USP2 expression decreased migration and invasion in LM2-4175 and SCP46 cells in association with the downregulation of matrix metalloproteinase-2 (MMP2) expression, whereas overexpression of USP2 in MDA-MB-468 and MDA-MB-231 cells enhanced migration and invasion and upregulated the expression of MMP2. The present study showed that USP2 expression is associated with TNBC cell line’s invasiveness and poor survival of breast cancer patients and may serve as a prognostic biomarker and therapeutic target for TNBC.


Triple negative breast cancer USP2 Metastasis MMP2 



We thank Dr. Guohong Hu from the Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, China, for providing LM2-4175, SCP2, SCP28, and SCP46 cell lines. Research in the authors’ laboratory is supported by the Scientific Research Project of Shanghai Municipal Health Bureau (20134Y039) and the National Natural Funding of China (81202087, 81172520, 81202088).

Conflicts of interest


Supplementary material

13277_2015_3207_MOESM1_ESM.docx (16 kb)
ESM 1 (DOCX 16 kb)


  1. 1.
    Perou CM, Sorlie T, Eisen MB, van de Rijn M, Jeffrey SS, Rees CA, et al. Molecular portraits of human breast tumours. Nature. 2000;406:747–52.CrossRefPubMedGoogle Scholar
  2. 2.
    Abd El-Rehim DM, Ball G, Pinder SE, Rakha E, Paish C, Robertson JF, et al. High-throughput protein expression analysis using tissue microarray technology of a large well-characterised series identifies biologically distinct classes of breast cancer confirming recent cDNA expression analyses. Int J Cancer J Int Cancer. 2005;116:340–50.CrossRefGoogle Scholar
  3. 3.
    Sorlie T, Tibshirani R, Parker J, Hastie T, Marron JS, Nobel A, et al. Repeated observation of breast tumor subtypes in independent gene expression data sets. Proc Natl Acad Sci U S A. 2003;100:8418–23.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Dent R, Trudeau M, Pritchard KI, Hanna WM, Kahn HK, Sawka CA, et al. Triple-negative breast cancer: clinical features and patterns of recurrence. Clin Cancer Res: Off J Am Assoc Cancer Res. 2007;13:4429–34.CrossRefGoogle Scholar
  5. 5.
    Tischkowitz M, Brunet JS, Begin LR, Huntsman DG, Cheang MC, Akslen LA, et al. Use of immunohistochemical markers can refine prognosis in triple negative breast cancer. BMC Cancer. 2007;7:134.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Bauer KR, Brown M, Cress RD, Parise CA, Caggiano V. Descriptive analysis of estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, and HER2-negative invasive breast cancer, the so-called triple-negative phenotype: a population-based study from the California Cancer Registry. Cancer. 2007;109:1721–8.CrossRefPubMedGoogle Scholar
  7. 7.
    Haffty BG, Yang Q, Reiss M, Kearney T, Higgins SA, Weidhaas J, et al. Locoregional relapse and distant metastasis in conservatively managed triple negative early-stage breast cancer. J Clin Oncol: Off J Am Soc Clin Oncol. 2006;24:5652–7.CrossRefGoogle Scholar
  8. 8.
    Eiermann W, Bergh J, Cardoso F, Conte P, Crown J, Curtin NJ, et al. Triple negative breast cancer: proposals for a pragmatic definition and implications for patient management and trial design. Breast. 2012;21:20–6.CrossRefPubMedGoogle Scholar
  9. 9.
    Lehmann BD, Bauer JA, Chen X, Sanders ME, Chakravarthy AB, Shyr Y, et al. Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. J Clin Invest. 2011;121:2750–67.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Amerik AY, Hochstrasser M. Mechanism and function of deubiquitinating enzymes. Biochim Biophys Acta. 2004;1695:189–207.CrossRefPubMedGoogle Scholar
  11. 11.
    Priolo C, Tang D, Brahamandan M, Benassi B, Sicinska E, Ogino S, et al. The isopeptidase USP2a protects human prostate cancer from apoptosis. Cancer Res. 2006;66:8625–32.CrossRefPubMedGoogle Scholar
  12. 12.
    Anders C, Carey LA. Understanding and treating triple-negative breast cancer. Oncology (Williston Park). 2008;22:1233–9. discussion 1239–1240, 1243.Google Scholar
  13. 13.
    Carey L, Winer E, Viale G, Cameron D, Gianni L. Triple-negative breast cancer: disease entity or title of convenience? Nat Rev Clin Oncol. 2010;7:683–92.CrossRefPubMedGoogle Scholar
  14. 14.
    Sacco JJ, Coulson JM, Clague MJ, Urbe S. Emerging roles of deubiquitinases in cancer-associated pathways. IUBMB Life. 2010;62:140–57.PubMedGoogle Scholar
  15. 15.
    Renatus M, Parrado SG, D’Arcy A, Eidhoff U, Gerhartz B, Hassiepen U, et al. Structural basis of ubiquitin recognition by the deubiquitinating protease USP2. Structure. 2006;14:1293–302.CrossRefPubMedGoogle Scholar
  16. 16.
    Nijman SM, Luna-Vargas MP, Velds A, Brummelkamp TR, Dirac AM, Sixma TK, et al. A genomic and functional inventory of deubiquitinating enzymes. Cell. 2005;123:773–86.CrossRefPubMedGoogle Scholar
  17. 17.
    Cummins JM, Vogelstein B. HAUSP is required for p53 destabilization. Cell Cycle. 2004;3:689–92.CrossRefPubMedGoogle Scholar
  18. 18.
    Li M, Brooks CL, Kon N, Gu W. A dynamic role of HAUSP in the p53-Mdm2 pathway. Mol Cell. 2004;13:879–86.CrossRefPubMedGoogle Scholar
  19. 19.
    Li M, Chen D, Shiloh A, Luo J, Nikolaev AY, Qin J, et al. Deubiquitination of p53 by HAUSP is an important pathway for p53 stabilization. Nature. 2002;416:648–53.CrossRefPubMedGoogle Scholar
  20. 20.
    Huang TT, Nijman SM, Mirchandani KD, Galardy PJ, Cohn MA, Haas W, et al. Regulation of monoubiquitinated PCNA by DUB autocleavage. Nat Cell Biol. 2006;8:339–47.PubMedGoogle Scholar
  21. 21.
    Gupta K, Copeland NG, Gilbert DJ, Jenkins NA, Gray DA. Unp, a mouse gene related to the tre oncogene. Oncogene. 1993;8:2307–10.PubMedGoogle Scholar
  22. 22.
    Frederick A, Rolfe M, Chiu MI. The human UNP locus at 3p21.31 encodes two tissue-selective, cytoplasmic isoforms with deubiquitinating activity that have reduced expression in small cell lung carcinoma cell lines. Oncogene. 1998;16:153–65.CrossRefPubMedGoogle Scholar
  23. 23.
    DeSalle LM, Latres E, Lin D, Graner E, Montagnoli A, Baker RT, et al. The de-ubiquitinating enzyme Unp interacts with the retinoblastoma protein. Oncogene. 2001;20:5538–42.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Qing Qu
    • 1
  • Yan Mao
    • 2
  • Gang Xiao
    • 3
  • Xiaochun Fei
    • 4
  • Jinglong Wang
    • 3
  • Yuzi Zhang
    • 2
  • Junjun Liu
    • 2
  • Guangcun Cheng
    • 3
  • Xiaosong Chen
    • 2
  • Jianhua Wang
    • 3
  • Kunwei Shen
    • 2
  1. 1.Department of Oncology, Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghaiChina
  2. 2.Comprehensive Breast Health Center, Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghaiChina
  3. 3.Department of Biochemistry and Molecular & Cell BiologyShanghai Jiao Tong University School of MedicineShanghaiChina
  4. 4.Department of Pathology, Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghaiChina

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