Identification of MET and SRC activation in melanoma cell lines showing primary resistance to PLX4032

Elisabetta Vergani, Viviana Vallacchi, Simona Frigerio, Paola Deho, Piera Mondellini, Paola Perego, Giuliana Cassinelli, Cinzia Lanzi, Maria Adele Testi, Licia Rivoltini, Italia Bongarzone, Monica Rodolfo

Research output: Contribution to journalArticle

Abstract

PLX4032/vemurafenib is a first-in-class small-molecule BRAFV600E inhibitor with clinical activity in patients with BRAF mutant melanoma. Nevertheless, drug resistance develops in treated patients, and strategies to overcome primary and acquired resistance are required. To explore the molecular mechanisms involved in primary resistance to PLX4032, we investigated its effects on cell proliferation and signaling in a panel of 27 genetically characterized patient-derived melanoma cell lines. Cell sensitivity to PLX4032 was dependent on BRAFV600E and independent from other gene alterations that commonly occur in melanoma such as PTEN loss, BRAF, and MITF gene amplification. Two cell lines lacking sensitivity to PLX4032 and harboring a different set of genetic alterations were studied as models of primary resistance. Treatment with the MEK inhibitor UO126 but not with PLX4032 inhibited cell growth and ERK activation. Resistance to PLX4032 was maintained after CRAF down-regulation by siRNA indicating alternative activation of MEK-ERK signaling. Genetic characterization by multiplex ligation-dependent probe amplification and analysis of phosphotyrosine signaling by MALDI-TOF mass spectrometry analysis revealed the activation of MET and SRC signaling, associated with the amplification of MET and of CTNNB1 and CCND1 genes, respectively. The combination of PLX4032 with drugs or siRNA targetingMET was effective in inhibiting cell growth and reducing cell invasion and migration in melanoma cells with MET amplification; similar effects were observed after targeting SRC in the other cell line, indicating a role for MET and SRC signaling in primary resistance to PLX4032. Our results support the development of classification of melanoma in molecular subtypes for more effective therapies.

Original languageEnglish
Pages (from-to)1132-1142
Number of pages11
JournalNeoplasia (United States)
Volume13
Issue number12
DOIs
Publication statusPublished - Dec 2011

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Melanoma
Cell Line
Mitogen-Activated Protein Kinase Kinases
Small Interfering RNA
Activation Analysis
PLX4032
Phosphotyrosine
Gene Amplification
Multiplex Polymerase Chain Reaction
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Growth
Drug Resistance
Genes
Cell Movement
Mass Spectrometry
Down-Regulation
Cell Proliferation
Therapeutics
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Cancer Research

Cite this

Identification of MET and SRC activation in melanoma cell lines showing primary resistance to PLX4032. / Vergani, Elisabetta; Vallacchi, Viviana; Frigerio, Simona; Deho, Paola; Mondellini, Piera; Perego, Paola; Cassinelli, Giuliana; Lanzi, Cinzia; Testi, Maria Adele; Rivoltini, Licia; Bongarzone, Italia; Rodolfo, Monica.

In: Neoplasia (United States), Vol. 13, No. 12, 12.2011, p. 1132-1142.

Research output: Contribution to journalArticle

Vergani, Elisabetta ; Vallacchi, Viviana ; Frigerio, Simona ; Deho, Paola ; Mondellini, Piera ; Perego, Paola ; Cassinelli, Giuliana ; Lanzi, Cinzia ; Testi, Maria Adele ; Rivoltini, Licia ; Bongarzone, Italia ; Rodolfo, Monica. / Identification of MET and SRC activation in melanoma cell lines showing primary resistance to PLX4032. In: Neoplasia (United States). 2011 ; Vol. 13, No. 12. pp. 1132-1142.
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