Immunohistochemical assessment of basal and luminal markers in non-muscle invasive urothelial carcinoma of bladder

  • Maria Del Carmen Rodriguez Pena
  • Alcides Chaux
  • Marie-Lisa Eich
  • Aline C. Tregnago
  • Diana Taheri
  • Walaa Borhan
  • Rajni Sharma
  • M. Katayoon Rezaei
  • George J. NettoEmail author
Original Article


The Cancer Genome Atlas project introduced genomic taxonomy of basal and luminal molecular subtypes in muscle invasive bladder cancer. Fewer studies have addressed the molecular classification in non-muscle invasive bladder cancer (NMIBC). Our aim is to assess the applicability of the proposed phenotypic classification for NMIBC. Three TMAs were constructed from 193 TURBT specimens of 60 bladder cancer patients treated at one of the authors’ institutions (1998–2008). Follow-up data on recurrence, grade, or stage progression was obtained. Immunohistochemistry was performed using an automated Ventana System for markers indicative of luminal (GATA3, CK20, ER, Uroplakin II, and HER2/neu) and basal (CK5/6 and CD44) phenotype. Marker expression was evaluated by 3 urologic pathologists. Using unadjusted logistic regression, we found significant association between tumor recurrence at next biopsy and CD44 expression (OR = 2.51, P = 0.03), tumor recurrence at any subsequent biopsy and ER expression (OR = 0.24, P = 0.04), and tumor grade progression at any subsequent biopsy and HER2/neu expression (OR = 0.24, P = 0.04). After adjusting for pathologic stage, we found a significant association between CK5/6 expression and tumor stage progression at either next or any subsequent biopsy (OR = 0.94, P = 0.006; and OR = 0.97, P = 0.02, respectively). Our findings suggest that individual immunohistochemical markers may be of value as prognostic factors in NMIBC.


Molecular classification Basal Luminal Immunohistochemistry Urothelial carcinoma Non-muscle invasive bladder cancer 


Author contributions

GJN conceived and designed the study and wrote, edited, and reviewed the manuscript. MCRP collected the data and wrote, edited, and reviewed the manuscript. AC and MLE analyzed the data and wrote, edited, and reviewed the manuscript. ACT collected the data and reviewed the manuscript. DT and WB collected the data and reviewed the manuscript. MKR collected samples and clinical data and reviewed the manuscript. RS performed the immunohistochemical analysis and reviewed the manuscript.


This study is supported in part by a grant from The Johns Hopkins Greenberg Bladder Cancer Institute and The Patana Fund of the Brady Urological Institute, Baltimore, MD.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

All authors gave final approval for publication. GJN takes full responsibility for the work as a whole, including the study design, access to data, and the decision to submit and publish the manuscript.

Informed consent

The Institutional Review Board approved this study.

Supplementary material

428_2019_2618_MOESM1_ESM.jpg (381 kb)
Supplementary Fig 1 A-F: Boxplots illustrating the correlation of marker expression and outcome. Marker score is depicted on y axis. Lighter box plots correspond to presence of event (e.g. recurrence or progression of tumor) while darker box plots correspond to absence of event. Labels on top of the boxplots correspond to adjusted (Bonferroni’s method) p values using Mann-Whitney’s U test (JPG 380 kb)
428_2019_2618_MOESM2_ESM.jpg (107 kb)
Supplementary Fig 2 A-F: Association of outcome measures with basal vs luminal dichotomous categorization of each tumor using CD44 and CK5/6 scores as surrogate for basal category and CK20 and Uroplakin II for luminal category. The highest score of any of the above 4 surrogate markers was applied to assign a basal vs luminal category in a given tumor (JPG 107 kb)
428_2019_2618_MOESM3_ESM.jpg (108 kb)
Supplementary Fig 3 A-F: Association of outcome measures with basal vs luminal dichotomous categorization of each tumor using CD44 and CK5/6 scores as surrogate for basal category and CK20 and Uroplakin II for luminal category. The higher value among the sum of the scores of the two markers in each category was applied to assign a basal vs luminal category in a given tumor (JPG 107 kb)
428_2019_2618_MOESM4_ESM.docx (14 kb)
ESM 4 (DOCX 14 kb)
428_2019_2618_MOESM5_ESM.docx (14 kb)
ESM 5 (DOCX 14 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Maria Del Carmen Rodriguez Pena
    • 1
  • Alcides Chaux
    • 2
  • Marie-Lisa Eich
    • 1
  • Aline C. Tregnago
    • 3
  • Diana Taheri
    • 4
  • Walaa Borhan
    • 5
  • Rajni Sharma
    • 3
  • M. Katayoon Rezaei
    • 6
  • George J. Netto
    • 1
    • 3
    Email author
  1. 1.Department of PathologyThe University of Alabama at BirminghamBirminghamUSA
  2. 2.Department of Scientific Research, School of Postgraduate StudiesNorte UniversityAsuncionParaguay
  3. 3.Department of PathologyJohns Hopkins UniversityBaltimoreUSA
  4. 4.Department of Pathology, Isfahan Kidney Diseases Research CenterIsfahan University of Medical SciencesIsfahanIran
  5. 5.Department of Pathology, College of MedicineTaibah UniversityMadinahSaudi Arabia
  6. 6.Department of PathologyGeorge Washington UniversityWashingtonUSA

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