Skip to main content

Advertisement

SpringerLink
Log in

Elevated expression of podocalyxin is associated with lymphatic invasion, basal-like phenotype, and clinical outcome in axillary lymph node-negative breast cancer

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

Abstract

Lymphatic invasion (LVI) is associated with disease recurrence in axillary node-negative (ANN) breast cancer. Using gene expression profiling of 105 ANN tumors, we found that podocalyxin (PODXL) was more highly expressed in tumors with LVI (LVI+) than in those without LVI (LVI−). Differences in PODXL expression were validated using real-time polymerase chain reaction as well as by immunohistochemistry in an independent set of 652 tumors on tissue microarrays. Disease-free survival (DFS) analyses were conducted for association of high PODXL protein expression with risk of distant recurrence overall and within breast cancer subtypes using both Cox and cure-rate models. High PODXL expression was associated with poor prognosis features including large tumor size, high histological grade, estrogen and progesterone receptor negativity, and with clinical alterations characteristic of the basal-like breast cancer phenotype. Surprisingly, despite having other poor prognosis characteristics, women with high PODXL expressing tumors had better long-term DFS in multivariate analysis with traditional clinicopathologic factors including LVI and HER2 status (P = 0.001). PODXL has the potential to be a useful biomarker for identifying good prognosis patients in characteristically poor prognosis breast cancer groups and may impact treatment of women with this disease.

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

Similar content being viewed by others

Abbreviations

ANN:

Axillary node-negative

CK5:

Cytokeratin 5

DFS:

Disease-free survival

EGFR:

Epidermal growth factor receptor

ER:

Estrogen receptor

GEA:

Gene expression array

HER2:

Human epidermal growth factor receptor 2

HR:

Hazard ratio

IHC:

Immunohistochemistry

K–M:

Kaplan–Meier

LVI:

Lymphatic invasion

OR:

Odds ratio

PH:

Proportional hazards

p53:

Protein 53

PODXL:

Podocalyxin

PgR:

Progesterone receptor

RT-PCR:

Real-time polymerase chain reaction

TMA:

Tissue microarray

References

  1. Bull SB, Ozcelik H, Pinnaduwage D, Blackstein ME, Sutherland DA, Pritchard KI, Tzontcheva AT, Sidlofsky S, Hanna WM, Qizilbash AH, Tweeddale ME, Fine S et al (2004) The combination of p53 mutation and neu/erbB-2 amplification is associated with poor survival in node-negative breast cancer. J Clin Oncol 22:86–96

    Article  PubMed  CAS  Google Scholar 

  2. Kerjaschki D, Poczewski H, Dekan G, Horvat R, Balzar E, Kraft N, Atkins RC (1986) Identification of a major sialoprotein in the glycocalyx of human visceral glomerular epithelial cells. J Clin Invest 78:1142–1149

    Article  PubMed  CAS  Google Scholar 

  3. Kershaw DB, Beck SG, Wharram BL, Wiggins JE, Goyal M, Thomas PE, Wiggins RC (1997) Molecular cloning and characterization of human podocalyxin-like protein. J Biol Chem 272:15708–15714

    Article  PubMed  CAS  Google Scholar 

  4. Sassetti C, Van Zante A, Rosen SD (2000) Identification of endoglycan, a member of the CD34/podocalyxin family of sialomucins. J Biol Chem 275:9001–9010

    Article  PubMed  CAS  Google Scholar 

  5. McNagny KM, Pettersson I, Rossi F, Flamme I, Shevchenko A, Mann M, Graf T (1997) Thrombomucin, a novel cell surface protein that defines thrombocytes and multipotent hematopoietic progenitors. J Cell Biol 138:1395–1407

    Article  PubMed  CAS  Google Scholar 

  6. Kerosuo L, Juvonen E, Alitalo R, Gylling M, Kerjaschki D, Miettinen A (2004) Podocalyxin in human haematopoietic cells. Br J Haematol 124:809–818

    Article  PubMed  CAS  Google Scholar 

  7. Doyonnas R, Kershaw DB, Duhme C, Merkens H, Chelliah S, Graf T, McNagny KM (2001) Anuria, omphalocele, and perinatal lethality in mice lacking the CD34-related protein podocalyxin. J Exp Med 194:13–27

    Article  PubMed  CAS  Google Scholar 

  8. Takeda T, Go WY, Orlando RA, Farquhar MG (2000) Expression of podocalyxin inhibits cell–cell adhesion and modifies junctional properties in Madin-Darby canine kidney cells. Mol Biol Cell 11:3219–3232

    PubMed  CAS  Google Scholar 

  9. Kerjaschki D, Vernillo AT, Farquhar MG (1985) Reduced sialyation of podocalyxin–the major sialoprotein of the rat kidney glomerulus–in aminonucleoside nephrosis. Am J Path 118:343–349

    PubMed  CAS  Google Scholar 

  10. Pericacho M, Alonso-Martin S, Larrucea S, González-Manchón C, Fernández D, Sanchez I, Ayuso MS, Parrilla R (2011) Diminished thrombogenic responses by deletion of the podocalyxin gene in mouse megakaryocytes. PLoS One 6:e26025

    Article  PubMed  CAS  Google Scholar 

  11. Alonso-Martin S, Nowakowski A, Larrucea S, Fernández D, Vilar-Egla M, Ayuso MS, Parrilla R (2010) Overexpression of podocalyxin in megakaryocytes and platelets decreases the bleeding time and enhances the agonist-induced aggregation of platelets. Thromb Res 125:e300–e305

    Article  PubMed  CAS  Google Scholar 

  12. Larsson A, Johansson ME, Wangefjord S, Gaber A, Nodin B, Kucharzewska P, Welinder C, Belting M, Eberhard J, Johnsson A, Uhlén M, Jirström K (2011) Overexpression of podocalyxin-like protein is an independent factor of poor prognosis in colorectal cancer. Br J Cancer 105:666–672

    Article  PubMed  CAS  Google Scholar 

  13. Somasiri A, Nielsen JS, Makretsov N, McCoy ML, Prentice L, Gilks CB, Chia SK, Gelmon KA, Kershaw DB, Huntsman DG, McNagny KM, Roskelley CD (2004) Overexpression of the anti-adhesin podocalyxin is an independent predictor of breast cancer progression. Cancer Res 64:5068–5073

    Article  PubMed  CAS  Google Scholar 

  14. Dallas MR, Chen SH, Streppel MM, Sharma S, Miatra A, Konstantopoulos K (2012) Sialofucosylated podocalyxin is a functional E- and L-selectin ligand expressed by metastatic pancreatic cancer cells. Am J Physiol 303:C616–C624

    Article  CAS  Google Scholar 

  15. Yasuoka H, Tsujimoto M, Inagaki M, Kodama R, Tsuji H, Iwahashi Y, Mabuchi Y, Ino K, Sanke T, Nakamura Y (2012) Clinicopathological significance of podocalyxin and phosphorylated ezrin in uterine endometroid adenocarcinoma. J Clin Pathol 65:399–402

    Article  PubMed  Google Scholar 

  16. Kelley TW, Huntsman D, McNagny KM, Roskelley CD, Hsi ED (2005) Podocalyxin: a marker of blasts in acute leukemia. Am J Clin Pathol 124:134–142

    Article  PubMed  Google Scholar 

  17. Heukamp LC, Fischer HP, Schirmacher P, Chen X, Breuhahn K, Nicolay C, Buttner R, Gutgemann I (2006) Podocalyxin-like protein 1 expression in primary hepatic tumours and tumour-like lesions. Histopathology 49:242–247

    Article  PubMed  CAS  Google Scholar 

  18. Naishiro Y, Yamada T, Idogawa M, Honda K, Takada M, Kondo T, Imai K, Hirohashi S (2005) Morphological and transcriptional responses of untransformed intestinal epithelial cells to an oncogenic beta-catenin protein. Oncogene 24:3141–3153

    Article  PubMed  CAS  Google Scholar 

  19. Barton Furness SG, McNagny K (2006) Beyond mere markers: functions for CD34 family of sialomucins in hematopoiesis. Immunol Res 34:13–32

    Article  Google Scholar 

  20. Larrucea S, Butta N, Arias-Salgado EG, Alonso-Martin S, Ayuso MS, Parrilla R (2008) Expression of podocalyxin enhances the adherence, migration, and intercellular communication of cells. Exp Cell Res 314:2004–2015

    Article  PubMed  CAS  Google Scholar 

  21. Sizemore S, Cicek M, Sizemore N, Ng KP, Casey G (2007) Podocalyxin increases the aggressive phenotype of breast and prostate cancer cells in vitro through its interaction with ezrin. Cancer Res 67:6183–6191

    Article  PubMed  CAS  Google Scholar 

  22. Sy JP, Taylor JM (2000) Estimation in a Cox proportional hazards cure model. Biometrics 56:227–236

    Article  PubMed  CAS  Google Scholar 

  23. Andrulis IL, Bull SB, Blackstein ME, Sutherland D, Mak C, Sidlofsky S, Pritzker KP, Hartwick RW, Hanna W, Lickley L, Wilkinson R, Qizilbash A et al (1998) neu/erbB-2 amplification identifies a poor-prognosis group of women with node-negative breast cancer. Toronto Breast Cancer Study Group. J Clin Oncol 16:1340–1349

    PubMed  CAS  Google Scholar 

  24. Mulligan AM, Pinnaduwage D, Bull SB, O’Malley FP, Andrulis IL (2008) Prognostic effect of basal-like breast cancers is time dependent: evidence from tissue microarray studies on a lymph node-negative cohort. Clin Cancer Res 14:4168–4174

    Article  PubMed  CAS  Google Scholar 

  25. McShane LM, Altman DG, Sauerbrei W, Taube SE, Gion M, Clark GM (2005) REporting recommendations for tumour MARKer prognostic studies (REMARK). Br J Cancer 93:387–391

    Article  PubMed  CAS  Google Scholar 

  26. Allred DC, Harvey JM, Berardo M, Clark GM (1998) Prognostic and predictive factors in breast cancer by immunohistochemical analysis. Mod Pathol 11:155–168

    PubMed  CAS  Google Scholar 

  27. Liu CL, Prapong W, Natkunam Y, Alizadeh A, Montgomery K, Gilks CB, van de Rijn M (2002) Software tools for high-throughput analysis and archiving of immunohistochemistry staining data obtained with tissue microarrays. Am J Pathol 161:1557–1565

    Article  PubMed  CAS  Google Scholar 

  28. Cox DR, Oakes D (1984) Analysis of survival data. Chapman and Hall, London

    Google Scholar 

  29. Farewell VT (1982) The use of mixture models for the analysis of survival data with long-term survivors. Biometrics 38:1041–1046

    Article  PubMed  CAS  Google Scholar 

  30. Heinze G, Ploner M (2002) SAS and SPLUS programs to perform Cox regression without convergence problems. Comput Methods Programs Biomed 67:217–223

    Article  PubMed  Google Scholar 

  31. Nielsen JS, McNagny KM (2009) The role of podocalyxin in health and disease. J Am Soc Nephrol 20:1669–1676

    Article  PubMed  CAS  Google Scholar 

  32. Harvey JM, Clark GM, Osborne CK, Allred DC (1999) Estrogen receptor status by immunohistochemistry is superior to the ligand-binding assay for predicting response to adjuvant endocrine therapy in breast cancer. J Clin Oncol 17:1474–1481

    PubMed  CAS  Google Scholar 

  33. Mohsin SK, Weiss H, Havighurst T, Clark GM, Berardo M, Roanh LD, To TV, Zho Q, Love RR, Allred DC (2004) Progesterone receptor by immunohistochemistry and clinical outcome in breast cancer: a validation study. Mod Pathol 17:1545–1554

    Article  PubMed  CAS  Google Scholar 

  34. O’ Malley FP, Parkes R, Latta E, Tjan S, Zadro T, Mueller R, Arneson N, Blackstein M, Andrulis I (2001) Comparison of HER2/neu status assessed by quantitative polymerase chain reaction and immunohistochemistry. Am J Clin Pathol 115:504–511

    Article  Google Scholar 

Download references

Acknowledgments

This study has been supported by a grant from the Canadian Institutes of Health Research and with funds provided by the Terry Fox Run (ILA, SBB, FO’M), by a Senior Investigator award from the Canadian Institutes of Health Research (SBB), and by project funds from the MITACS Network of Centres of Excellence in Mathematical Sciences (SBB). YEY is supported by MITACS Industrial Fellowship and a CIHR Fellow in Genetic Epidemiology and Statistical Genetics with CIHR STAGE (Strategic Training for Advanced Genetic Epidemiology). The authors thank the study participants and gratefully acknowledge the technical assistance of Lucine Bosnoyan-Collins and Suzanne Tjan and the contributions of the Toronto Breast Cancer Group to this work. We also thank Candemir Cigsar for statistical insights.

Conflict of interest

The authors declare they have no conflicts of interest.

Author information

Authors and Affiliations

  1. Fred A. Litwin Centre for Cancer Genetics, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, ON, M5G 1X5, Canada

    Catherine L. Forse & Irene L. Andrulis

  2. Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 60 Murray Street, Toronto, ON, M5T 3L9, Canada

    Yildiz E. Yilmaz, Dushanthi Pinnaduwage & Shelley B. Bull

  3. Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada

    Catherine L. Forse & Irene L. Andrulis

  4. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada

    Frances P. O’Malley, Anna Marie Mulligan & Irene L. Andrulis

  5. Dalla Lana School of Public Health, University of Toronto, Toronto, ON, Canada

    Yildiz E. Yilmaz & Shelley B. Bull

  6. Department of Laboratory Medicine and the Keenan Research Centre of the Li Ka Shing Knowledge Institute, St. Michael’s Hospital, Toronto, ON, Canada

    Frances P. O’Malley

  7. Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, ON, Canada

    Irene L. Andrulis

  8. Department of Pathology, University Health Network, Toronto, ON, Canada

    Anna Marie Mulligan

Authors
  1. Catherine L. Forse
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yildiz E. Yilmaz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dushanthi Pinnaduwage
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Frances P. O’Malley
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Anna Marie Mulligan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shelley B. Bull
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Irene L. Andrulis
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Irene L. Andrulis.

Additional information

C. L. Forse and Y. E. Yilmaz contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Forse, C.L., Yilmaz, Y.E., Pinnaduwage, D. et al. Elevated expression of podocalyxin is associated with lymphatic invasion, basal-like phenotype, and clinical outcome in axillary lymph node-negative breast cancer. Breast Cancer Res Treat 137, 709–719 (2013). https://doi.org/10.1007/s10549-012-2392-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-012-2392-y

Keywords

Search

Navigation