Proteomics study of medullary thyroid carcinomas expressing RET germ-line mutations: Identification of new signaling elements

L. Gorla, P. Mondellini, G. Cuccuru, F. Miccichè, G. Cassinelli, M. Cremona, M. A. Pierotti, C. Lanzi, I. Bongarzone

Research output: Contribution to journalArticle

Abstract

Proteomics may help to elucidate differential signaling networks underlying the effects of compounds and to identify new therapeutic targets. Using a proteomic-multiplexed analysis of the phosphotyrosine signaling together with antibody-based validation techniques, we identified several candidate molecules for RET (rearranged during transfection) tyrosine kinase receptor carrying mutations responsible for the multiple endocrine neoplasia type 2A and 2B (MEN2A and MEN2B) syndromes in two human medullary thyroid carcinoma (MTC) cell lines, TT and MZ-CRC-1, which express the RET-MEN2A and RET-MEN2B oncoproteins, respectively. Signaling elements downstream of these oncoproteins were identified after treating cells with the indolinone tyrosine kinase inhibitor RPI-1 to knock down RET phosphorylation activity. We detected 23 and 18 affinity-purified phosphotyrosine proteins in untreated TT and MZ-CRC-1 cells, respectively, most of which were shared and sensitive to RPI-1 treatment. However, our data clearly point to specific signaling features of the RET-MEN2A and RET-MEN2B oncogenic pathways. Moreover, the detection of high-level expression of minimally phosphorylated epidermal growth factor receptor (EGFR) in both TT and MZ-CRC-1 cells, together with our data on the effects of EGF stimulation on the proteomic profiles and the response to Gefitinib treatment, suggest the relevance of EGFR signaling in these cell lines, especially since analysis of 14 archival MTC specimens revealed EGFR mRNA expression in all samples. Together, our data suggest that RET/EGFR multi-target inhibitors might be beneficial for therapy of MTC.

Original languageEnglish
Pages (from-to)220-231
Number of pages12
JournalMolecular Carcinogenesis
Volume48
Issue number3
DOIs
Publication statusPublished - Mar 2009

Fingerprint

Germ-Line Mutation
Proteomics
Transfection
Epidermal Growth Factor Receptor
Phosphotyrosine
Oncogene Proteins
Multiple Endocrine Neoplasia Type 2b
Multiple Endocrine Neoplasia Type 2a
Cell Line
Receptor Protein-Tyrosine Kinases
Therapeutics
Medullary Thyroid cancer
Epidermal Growth Factor
Protein-Tyrosine Kinases
Phosphorylation
Messenger RNA
Mutation
Antibodies

Keywords

  • EGFR
  • LGALS3BP
  • Medullary thyroid carcinoma
  • RET oncogenic signaling
  • Tyrosine phosphorylation

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology

Cite this

Proteomics study of medullary thyroid carcinomas expressing RET germ-line mutations : Identification of new signaling elements. / Gorla, L.; Mondellini, P.; Cuccuru, G.; Miccichè, F.; Cassinelli, G.; Cremona, M.; Pierotti, M. A.; Lanzi, C.; Bongarzone, I.

In: Molecular Carcinogenesis, Vol. 48, No. 3, 03.2009, p. 220-231.

Research output: Contribution to journalArticle

Gorla, L. ; Mondellini, P. ; Cuccuru, G. ; Miccichè, F. ; Cassinelli, G. ; Cremona, M. ; Pierotti, M. A. ; Lanzi, C. ; Bongarzone, I. / Proteomics study of medullary thyroid carcinomas expressing RET germ-line mutations : Identification of new signaling elements. In: Molecular Carcinogenesis. 2009 ; Vol. 48, No. 3. pp. 220-231.
@article{7b5c82210a5d4c9fa44df28954f80496,
title = "Proteomics study of medullary thyroid carcinomas expressing RET germ-line mutations: Identification of new signaling elements",
abstract = "Proteomics may help to elucidate differential signaling networks underlying the effects of compounds and to identify new therapeutic targets. Using a proteomic-multiplexed analysis of the phosphotyrosine signaling together with antibody-based validation techniques, we identified several candidate molecules for RET (rearranged during transfection) tyrosine kinase receptor carrying mutations responsible for the multiple endocrine neoplasia type 2A and 2B (MEN2A and MEN2B) syndromes in two human medullary thyroid carcinoma (MTC) cell lines, TT and MZ-CRC-1, which express the RET-MEN2A and RET-MEN2B oncoproteins, respectively. Signaling elements downstream of these oncoproteins were identified after treating cells with the indolinone tyrosine kinase inhibitor RPI-1 to knock down RET phosphorylation activity. We detected 23 and 18 affinity-purified phosphotyrosine proteins in untreated TT and MZ-CRC-1 cells, respectively, most of which were shared and sensitive to RPI-1 treatment. However, our data clearly point to specific signaling features of the RET-MEN2A and RET-MEN2B oncogenic pathways. Moreover, the detection of high-level expression of minimally phosphorylated epidermal growth factor receptor (EGFR) in both TT and MZ-CRC-1 cells, together with our data on the effects of EGF stimulation on the proteomic profiles and the response to Gefitinib treatment, suggest the relevance of EGFR signaling in these cell lines, especially since analysis of 14 archival MTC specimens revealed EGFR mRNA expression in all samples. Together, our data suggest that RET/EGFR multi-target inhibitors might be beneficial for therapy of MTC.",
keywords = "EGFR, LGALS3BP, Medullary thyroid carcinoma, RET oncogenic signaling, Tyrosine phosphorylation",
author = "L. Gorla and P. Mondellini and G. Cuccuru and F. Miccich{\`e} and G. Cassinelli and M. Cremona and Pierotti, {M. A.} and C. Lanzi and I. Bongarzone",
year = "2009",
month = "3",
doi = "10.1002/mc.20474",
language = "English",
volume = "48",
pages = "220--231",
journal = "Molecular Carcinogenesis",
issn = "0899-1987",
publisher = "Wiley-Liss Inc.",
number = "3",

}

TY - JOUR

T1 - Proteomics study of medullary thyroid carcinomas expressing RET germ-line mutations

T2 - Identification of new signaling elements

AU - Gorla, L.

AU - Mondellini, P.

AU - Cuccuru, G.

AU - Miccichè, F.

AU - Cassinelli, G.

AU - Cremona, M.

AU - Pierotti, M. A.

AU - Lanzi, C.

AU - Bongarzone, I.

PY - 2009/3

Y1 - 2009/3

N2 - Proteomics may help to elucidate differential signaling networks underlying the effects of compounds and to identify new therapeutic targets. Using a proteomic-multiplexed analysis of the phosphotyrosine signaling together with antibody-based validation techniques, we identified several candidate molecules for RET (rearranged during transfection) tyrosine kinase receptor carrying mutations responsible for the multiple endocrine neoplasia type 2A and 2B (MEN2A and MEN2B) syndromes in two human medullary thyroid carcinoma (MTC) cell lines, TT and MZ-CRC-1, which express the RET-MEN2A and RET-MEN2B oncoproteins, respectively. Signaling elements downstream of these oncoproteins were identified after treating cells with the indolinone tyrosine kinase inhibitor RPI-1 to knock down RET phosphorylation activity. We detected 23 and 18 affinity-purified phosphotyrosine proteins in untreated TT and MZ-CRC-1 cells, respectively, most of which were shared and sensitive to RPI-1 treatment. However, our data clearly point to specific signaling features of the RET-MEN2A and RET-MEN2B oncogenic pathways. Moreover, the detection of high-level expression of minimally phosphorylated epidermal growth factor receptor (EGFR) in both TT and MZ-CRC-1 cells, together with our data on the effects of EGF stimulation on the proteomic profiles and the response to Gefitinib treatment, suggest the relevance of EGFR signaling in these cell lines, especially since analysis of 14 archival MTC specimens revealed EGFR mRNA expression in all samples. Together, our data suggest that RET/EGFR multi-target inhibitors might be beneficial for therapy of MTC.

AB - Proteomics may help to elucidate differential signaling networks underlying the effects of compounds and to identify new therapeutic targets. Using a proteomic-multiplexed analysis of the phosphotyrosine signaling together with antibody-based validation techniques, we identified several candidate molecules for RET (rearranged during transfection) tyrosine kinase receptor carrying mutations responsible for the multiple endocrine neoplasia type 2A and 2B (MEN2A and MEN2B) syndromes in two human medullary thyroid carcinoma (MTC) cell lines, TT and MZ-CRC-1, which express the RET-MEN2A and RET-MEN2B oncoproteins, respectively. Signaling elements downstream of these oncoproteins were identified after treating cells with the indolinone tyrosine kinase inhibitor RPI-1 to knock down RET phosphorylation activity. We detected 23 and 18 affinity-purified phosphotyrosine proteins in untreated TT and MZ-CRC-1 cells, respectively, most of which were shared and sensitive to RPI-1 treatment. However, our data clearly point to specific signaling features of the RET-MEN2A and RET-MEN2B oncogenic pathways. Moreover, the detection of high-level expression of minimally phosphorylated epidermal growth factor receptor (EGFR) in both TT and MZ-CRC-1 cells, together with our data on the effects of EGF stimulation on the proteomic profiles and the response to Gefitinib treatment, suggest the relevance of EGFR signaling in these cell lines, especially since analysis of 14 archival MTC specimens revealed EGFR mRNA expression in all samples. Together, our data suggest that RET/EGFR multi-target inhibitors might be beneficial for therapy of MTC.

KW - EGFR

KW - LGALS3BP

KW - Medullary thyroid carcinoma

KW - RET oncogenic signaling

KW - Tyrosine phosphorylation

UR - http://www.scopus.com/inward/record.url?scp=62349122222&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=62349122222&partnerID=8YFLogxK

U2 - 10.1002/mc.20474

DO - 10.1002/mc.20474

M3 - Article

C2 - 18756447

AN - SCOPUS:62349122222

VL - 48

SP - 220

EP - 231

JO - Molecular Carcinogenesis

JF - Molecular Carcinogenesis

SN - 0899-1987

IS - 3

ER -