Cellular effects and antitumor activity of RET inhibitor RPI-1 on MEN2A-associated medullary thyroid carcinoma

Giuditta Cuccuru, Cinzia Lanzi, Giuliana Cassinelli, Graziella Pratesi, Monica Tortoreto, Giovanna Petrangolini, Ettore Seregni, Antonia Martinetti, Diletta Laccabue, Chiara Zanchi, Franco Zunino

Research output: Contribution to journalArticle

Abstract

Background: The RET proto-oncogene encodes a receptor tyrosine kinase. RET oncogenes arise through sporadic and inherited gene mutations and are involved in the etiopathogenesis of medullary thyroid carcinoma, a cancer that responds poorly to conventional chemotherapy. Medullary thyroid carcinoma is a component of multiple endocrine neoplasia type 2 or MEN2 syndromes. Methods: We investigated the cellular effects of RPI-1, a novel 2-indolinone Ret tyrosine kinase inhibitor on cells that express RET C634 oncogenic mutants common in the MEN2A syndrome: NIH3T3 fibroblasts transfected with RETC634R and human medullary thyroid carcinoma TT cells that express endogenous RETC634W. RPI-1 antiproliferative activity was determined by cell proliferation and anchorage-independent growth assays. Expression and phosphorylation of Ret and of proteins involved in downstream signaling pathways were examined by immunoblotting. Antitumor activity of oral RPI-1 treatment was tested by using two dosing levels in nude mice bearing subcutaneous TT xenograft tumors. All statistical tests were two-sided. Results: The RPI-1 IC50 value for cell proliferation was 3.6 μM (95% confidence interval [CI] = 1.8 to 5.4 μM) in NIH3T3 cells expressing the Ret mutant compared with 16 μM (95% CI = 12.3 to 19.7 μM) in non-transfected NIH3T3 cells, and that for colony formation in soft agar was 2.4 μM (95% CI = 0.8 to 4.0 μM) and 26 μM (95% CI = 17 to 35 μM) in RET mutant-transfected and H-RAS-transfected NIH3T3 cells, respectively. In NIH3T3 cells expressing the Ret mutant, Ret protein and tyrosine phosphorylation were undetectable after 24 hours of RPI-1 treatment. In TT cells, RPI-1 inhibited proliferation, Ret tyrosine phosphorylation, Ret protein expression, and the activation of PLCγ, ERKs and AKT. In mice, oral daily RPI-1 treatment inhibited the tumor growth of TT xenografts by 81% (P

Original languageEnglish
Pages (from-to)1006-1014
Number of pages9
JournalJournal of the National Cancer Institute
Volume96
Issue number13
Publication statusPublished - Jul 7 2004

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Confidence Intervals
Phosphorylation
Heterografts
Tyrosine
Cell Proliferation
Multiple Endocrine Neoplasia Type 2a
Neoplasms
Proto-Oncogenes
Receptor Protein-Tyrosine Kinases
Mutant Proteins
Growth
Medullary Thyroid cancer
Oncogenes
Immunoblotting
Nude Mice
Protein-Tyrosine Kinases
Inhibitory Concentration 50
Agar
Proteins
Therapeutics

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Cuccuru, G., Lanzi, C., Cassinelli, G., Pratesi, G., Tortoreto, M., Petrangolini, G., ... Zunino, F. (2004). Cellular effects and antitumor activity of RET inhibitor RPI-1 on MEN2A-associated medullary thyroid carcinoma. Journal of the National Cancer Institute, 96(13), 1006-1014.

Cellular effects and antitumor activity of RET inhibitor RPI-1 on MEN2A-associated medullary thyroid carcinoma. / Cuccuru, Giuditta; Lanzi, Cinzia; Cassinelli, Giuliana; Pratesi, Graziella; Tortoreto, Monica; Petrangolini, Giovanna; Seregni, Ettore; Martinetti, Antonia; Laccabue, Diletta; Zanchi, Chiara; Zunino, Franco.

In: Journal of the National Cancer Institute, Vol. 96, No. 13, 07.07.2004, p. 1006-1014.

Research output: Contribution to journalArticle

Cuccuru, G, Lanzi, C, Cassinelli, G, Pratesi, G, Tortoreto, M, Petrangolini, G, Seregni, E, Martinetti, A, Laccabue, D, Zanchi, C & Zunino, F 2004, 'Cellular effects and antitumor activity of RET inhibitor RPI-1 on MEN2A-associated medullary thyroid carcinoma', Journal of the National Cancer Institute, vol. 96, no. 13, pp. 1006-1014.
Cuccuru, Giuditta ; Lanzi, Cinzia ; Cassinelli, Giuliana ; Pratesi, Graziella ; Tortoreto, Monica ; Petrangolini, Giovanna ; Seregni, Ettore ; Martinetti, Antonia ; Laccabue, Diletta ; Zanchi, Chiara ; Zunino, Franco. / Cellular effects and antitumor activity of RET inhibitor RPI-1 on MEN2A-associated medullary thyroid carcinoma. In: Journal of the National Cancer Institute. 2004 ; Vol. 96, No. 13. pp. 1006-1014.
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AU - Lanzi, Cinzia

AU - Cassinelli, Giuliana

AU - Pratesi, Graziella

AU - Tortoreto, Monica

AU - Petrangolini, Giovanna

AU - Seregni, Ettore

AU - Martinetti, Antonia

AU - Laccabue, Diletta

AU - Zanchi, Chiara

AU - Zunino, Franco

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