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Mutational status of VHL gene and its clinical importance in renal clear cell carcinoma

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Abstract

The most common subtype of renal cell carcinoma is the clear cell type (ccRCC), accounting for 75 % of cases. Inactivation of VHL gene is thought to be an early event in ccRCC carcinogenesis. Our intention was to assess whether VHL mutational status might provide useful predictive or prognostic information in patients with ccRCC. VHL messenger RNA (mRNA) expression was analyzed by in situ hybridization and its protein by immunohistochemistry on a tissue microarray containing samples from 148 cases. This was validated by qRT-PCR on 62 cases, for which RNA was available. The mutation status was assessed in 91 cases by Sanger sequencing. VHL was found mutated in 57 % of cases, with missense mutations in 26 %, nonsense in 5 %, splice site in 13 %, deletions in 39 %, indels in 8 %, duplications in 8 %, and insertions in 2 % of the cases. The prevalence of mutations by exon was the following: exon 1, 47 %; exon 2, 27 %; and exon 3, 13 %. VHL protein was expressed in a high number of cases (80 %), but significant correlations were not found between protein expression, clinical data, and survival. Importantly, of the 91 samples evaluated by sequencing, 45 were mutated, and 87 % of those were strongly positive. We found 32 novel mutations in the VHL gene in ccRCC. The presence of mutations was not concordant with mRNA or protein expression. Nonsense mutations of the VHL gene appear to be related with poorer prognosis and survival.

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Abbreviations

ccRCC:

Clear cel renal carcinoma

Cul2:

Cullin 2

FFPE:

Formalin-fixed paraffin-embedded

HGMD:

The Human Gene Mutation Database

HIF1α:

Hypoxia-inducible factor 1-alpha

IHC:

Immunohistochemistry

ISH:

In situ hybridization

Rbx-1:

Ring-box protein 1

rcc:

Renal cell carcinoma

RT-PCR:

Reverse transcription polymerase chain reaction

SAME:

Department of Medical Records and Statistics

TNM:

Classification of malignant Tumors

VHL:

Von Hippel–Lindau

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Acknowledgments

This work was funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP).

Conflict of interest

The authors declare that they have no competing interests.

Funding

This work was funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP).

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Authors and Affiliations

  1. International Center of Research in Cancer (CIPE), A.C. Camargo Cancer Center, Rua Taguá, 440-Liberdade, 01508-010, Sao Paulo, SP, Brazil

    Mariana Rezende Alves, Felipe Cavalcanti Carneiro, André Mourão Lavorato-Rocha, Dirce Maria Carraro & Rafael Malagoli Rocha

  2. Urology Division Department, A.C. Camargo Cancer Center, Rua Professor Antônio Prudente, 211 - Liberdade, 01509-010, São Paulo, SP, Brazil

    Walter Henriques da Costa, Stênio de Cássio Zequi & Gustavo Cardoso Guimaraes

  3. Department of Investigative Pathology, A.C. Camargo Cancer Center, Rua Professor Antônio Prudente, 211 - Liberdade, 01509-010, São Paulo, SP, Brazil

    Isabela Werneck da Cunha, Fernando Augusto Soares & Rafael Malagoli Rocha

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  1. Mariana Rezende Alves
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  2. Felipe Cavalcanti Carneiro
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Correspondence to Rafael Malagoli Rocha.

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Alves, M.R., Carneiro, F.C., Lavorato-Rocha, A.M. et al. Mutational status of VHL gene and its clinical importance in renal clear cell carcinoma. Virchows Arch 465, 321–330 (2014). https://doi.org/10.1007/s00428-014-1629-z

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  • DOI: https://doi.org/10.1007/s00428-014-1629-z

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