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Identification and characterization of two novel germline RET variants associated with medullary thyroid carcinoma

  • Endocrine Genetics/Epigenetics
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Abstract

Activating germline mutations in the RET proto-oncogene are responsible for about 98 % of the familial forms of medullary thyroid carcinoma (MTC), which represent 25 % of all MTC cases. The search for germline mutations in this gene is important for the recognition of hereditary forms of MTC and further identification of at-risk relatives who may benefit from early clinical intervention. Genotype–phenotype correlations are well established for most disease-causing RET mutations, allowing risk stratification. The association of a new RET variant with the MTC phenotype and familial predisposition requires the assessment of its functional and clinical significance. The aim of this study was to evaluate the oncogenic potential of two newly identified RET germline variants associated with late-onset MTC. In vitro functional assays were designed to address the transforming potential of novel RET variants, through their expression in non-transformed cells, and comparing their effect with wild-type RET. The new variants were identified in codons 515 (p.C515W) and 636 (p.T636M) located, respectively, in exons 8 and 11, thus resulting in amino acid substitutions in the extracellular region of the tyrosine kinase receptor RET. Through functional assays, we observed increased cell growth and proliferation, loss of contact inhibition, and a stimulation of cell migration, suggesting that these new RET variants hold some relevant transforming potential. The transforming potential of these novel RET variants was of low-grade, when compared to that of RET MEN2A-causing mutation p.C634R, probably explaining the mild phenotype characterized by late onset and low clinical aggressiveness.

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Acknowledgments

We thank Dr Diogo Freitas dos Santos for referring us patient A. Moreover, we thank Dr. Masahide Takahashi for kindly donating pcDNA3.1/Zeo(+) plasmid carrying the human wild-type RET cDNA sequence. We also thank Paulo Matos (FCUL, Lisboa) as well as Bruno Cardoso (IPOFG, Lisboa) for generously providing pRK5-Myc-H-RASV12-expressing construct, pERK1/2 and pS6 antibodies.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Unidade de Investigação de Patobiologia Molecular, Instituto Português de Oncologia de Lisboa Francisco Gentil E.P.E., Rua Professor Lima Basto, 1099-023, Lisbon, Portugal

    A. L. Silva, F. Carmo, M. M. Moura, R. Domingues, C. Espadinha, V. Leite, B. Cavaco & M. J. Bugalho

  2. Serviço de Endocrinologia, Instituto Português de Oncologia de Lisboa Francisco Gentil E.P.E., 1099-023, Lisbon, Portugal

    V. Leite & M. J. Bugalho

  3. iNOVA4Health - Programme in Translational Medicine (iBET, CEDOC/NMS, IPOLFG and ITQB), Av. República , Qta. do Marquês, 2780-157, Oeiras, Portugal

    A. L. Silva, M. M. Moura, R. Domingues, V. Leite, B. Cavaco & M. J. Bugalho

  4. NOVA Medical School/Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Campo dos Mártires da Pátria, 130, 1169-056, Lisbon, Portugal

    V. Leite, B. Cavaco & M. J. Bugalho

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  1. A. L. Silva
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  2. F. Carmo
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  3. M. M. Moura
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  7. B. Cavaco
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  8. M. J. Bugalho
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Correspondence to A. L. Silva.

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Silva, A.L., Carmo, F., Moura, M.M. et al. Identification and characterization of two novel germline RET variants associated with medullary thyroid carcinoma. Endocrine 49, 366–372 (2015). https://doi.org/10.1007/s12020-015-0559-0

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  • DOI: https://doi.org/10.1007/s12020-015-0559-0

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