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Expression of CCN3 protein in human Wilms’ tumors: immunohistochemical detection of CCN3 variants using domain-specific antibodies

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Abstract

We aimed to detect truncated CCN3 protein variants in formalin-fixed paraffin-embedded samples of eight Wilms’ tumors using anti-K19M and novel domain-specific antibodies, anti-NH2, anti-NH3, anti-NH4, and anti-NH5 raised against C-terminal (CT) domain and modules 1, 2, 3, and 4 of the CCN3 protein, respectively. In Wilms’ tumors, all the domain antibodies except anti-NH4 exhibited both nuclear and cytoplasmic staining in blastema as well as primitive tubules. NH4 was detected only in the cytoplasm of tumor cells. Normal fetal kidneys revealed mainly cytoplasmic immunoreactivity for all antibodies in tubules and glomeruli, except for K19 and NH5, which showed some nuclear staining. Our data suggest expression of a truncated nuclear CCN3 variant lacking the thrombospondin type-1-like domain and cytoplasmic full-length CCN3 protein in Wilms’ tumor cells. In addition, normal fetal kidneys express mainly full-length protein mostly localized to cytoplasm. Truncated CCN3 protein in Wilms’ tumor cells may provide evidence for its tumorigenic role in these tumors. Uniform NH5 staining compared to variable expression of K19M indicates that using NH5 is a better approach for detecting the CT domain of CCN3 protein in archival specimens. Thus, the domain-specific antibodies represent valuable tools for detecting CCN3 protein variants in normal and neoplastic kidneys.

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Abbreviations

NOV:

Nephroblastoma overexpressed gene

MAV-1:

Myeloblastosis-associated virus type 1

SIOP:

International Society of Pediatric Oncology

CTGF:

Connective tissue growth factor

IGFBP:

Insulin-like growth factor binding protein

VWC:

Von Willebrand complex

CT:

C-terminal domain

TSP1:

Thrombospondin type 1 repeat

WT1:

Wilms’ tumor gene 1

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Acknowledgment

This work was supported by a grant (PI-04/0822) from the FIS (Instituto Carlos-III, Madrid, Spain) and PROTHETS (Prognosis and therapeutic targets of Ewing’s family of tumors) FP6 contract number 503036. Manish Mani Subramaniam is a holder of a grant from the Hospital Clinic of University and the Spanish Association Against Cancer (AECC) of Valencia, Spain.

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Authors and Affiliations

  1. Department of Pathology, Medical School, University of Valencia, Avda. Blasco Ibañez, 17, 46010, Valencia, Spain

    Manish Mani Subramaniam, Samuel Navarro & Antonio Llombart-Bosch

  2. Laboratoire d’ Oncologie Virale et Moléculaire, UFR de Biochimie, Université Paris 7-D.Diderot, Paris, France

    Noureddine Lazar & Bernard Perbal

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  1. Manish Mani Subramaniam
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  2. Noureddine Lazar
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  3. Samuel Navarro
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  4. Bernard Perbal
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  5. Antonio Llombart-Bosch
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Correspondence to Antonio Llombart-Bosch.

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Manish Mani Subramaniam and Noureddine Lazar made an equal contribution to this work.

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Subramaniam, M.M., Lazar, N., Navarro, S. et al. Expression of CCN3 protein in human Wilms’ tumors: immunohistochemical detection of CCN3 variants using domain-specific antibodies. Virchows Arch 452, 33–39 (2008). https://doi.org/10.1007/s00428-007-0523-3

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  • DOI: https://doi.org/10.1007/s00428-007-0523-3

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