Association of activated c‐Met with NRAS‐mutated human melanomas

Cutaneous melanomas can be divided into three mutually exclusive genetic subsets: tumors with mutated BRAF, tumors with mutated NRAS and tumors wild type at both loci (wt/wt). Targeted therapy for melanoma has been advancing with agents directed to mutated BRAF, accounting for 50% of melanoma patients. The c‐Met pathway is known to play a role in melanoma tumorigenesis and preliminary data from our laboratory suggested that this pathway is preferentially activated in NRAS‐mutated tumors. The objective of this study was to test the hypothesis that melanomas carrying the mutated NRAS genotype are uniquely sensitively to c‐Met inhibition, thus providing rationale for therapeutic targeting of c‐Met in this patient cohort. Using primary human melanomas with known BRAF/NRAS genotypes, we observed greater immunostaining for phosphorylated (activated) c‐Met in NRAS‐mutated and wt/wt tumors, compared to BRAF‐mutated tumors. NRAS‐mutated and wt/wt cell lines also demonstrated more robust c‐Met activation in response to hepatocyte growth factor (HGF). Knock‐down of mutated N‐Ras, but not wild type N‐Ras, by RNA interference resulted in decreased c‐Met phosphorylation. Compared to BRAF mutants, NRAS‐mutated melanoma cells were more sensitive to pharmacologic c‐Met inhibition in terms of c‐Met activation, Akt phosphorylation, tumor cell proliferation, migration and apoptosis. This enhanced sensitivity was observed in wt/wt cells as well, but was a less consistent finding. On the basis of these experimental results, we propose that c‐Met inhibition may be a useful therapeutic strategy for melanomas with NRAS mutations, as well as some tumors with a wt/wt genotype.

[1]  A. Hauschild,et al.  Improved survival with vemurafenib in melanoma with BRAF V600E mutation. , 2011, The New England journal of medicine.

[2]  Li-E. Wang,et al.  Clinical Correlates of NRAS and BRAF Mutations in Primary Human Melanoma , 2010, Clinical Cancer Research.

[3]  Y. Samuels,et al.  Analysis of the genome to personalize therapy for melanoma , 2010, Oncogene.

[4]  S. Ishikawa,et al.  Molecular Predictors of Sensitivity to the MET Inhibitor PHA665752 in Lung Carcinoma Cells , 2010, Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer.

[5]  Stephen K Burley,et al.  SGX523 is an exquisitely selective, ATP-competitive inhibitor of the MET receptor tyrosine kinase with antitumor activity in vivo , 2009, Molecular Cancer Therapeutics.

[6]  Paolo Romano,et al.  Cell Line Data Base: structure and recent improvements towards molecular authentication of human cell lines , 2008, Nucleic Acids Res..

[7]  Gary L Clayman,et al.  Small molecule c‐MET inhibitor PHA665752: Effect on cell growth and motility in papillary thyroid carcinoma , 2008, Head & neck.

[8]  S. Eschrich,et al.  The gene expression profiles of primary and metastatic melanoma yields a transition point of tumor progression and metastasis , 2008, BMC Medical Genomics.

[9]  D. Bottaro,et al.  Targeting the c-Met signaling pathway in cancer. , 2008, Clinical cancer research : an official journal of the American Association for Cancer Research.

[10]  Shinji Yamazaki,et al.  Cytoreductive antitumor activity of PF-2341066, a novel inhibitor of anaplastic lymphoma kinase and c-Met, in experimental models of anaplastic large-cell lymphoma , 2007, Molecular Cancer Therapeutics.

[11]  I. Kasman,et al.  MetMAb, the one-armed 5D5 anti-c-Met antibody, inhibits orthotopic pancreatic tumor growth and improves survival. , 2007, Cancer research.

[12]  R. Salgia,et al.  Downstream signalling and specific inhibition of c-MET/HGF pathway in small cell lung cancer: implications for tumour invasion , 2007, British Journal of Cancer.

[13]  Shinji Yamazaki,et al.  An orally available small-molecule inhibitor of c-Met, PF-2341066, exhibits cytoreductive antitumor efficacy through antiproliferative and antiangiogenic mechanisms. , 2007, Cancer research.

[14]  R. Salgia,et al.  c-Met Is a Potentially New Therapeutic Target for Treatment of Human Melanoma , 2007, Clinical Cancer Research.

[15]  Jilin Sun,et al.  Fully human monoclonal antibodies to hepatocyte growth factor with therapeutic potential against hepatocyte growth factor/c-Met-dependent human tumors. , 2006, Cancer research.

[16]  W. Shih,et al.  Detection of B-RAF and N-RAS mutations in human melanoma. , 2005, Journal of the American College of Surgeons.

[17]  Ravi Salgia,et al.  Functional expression and mutations of c-Met and its therapeutic inhibition with SU11274 and small interfering RNA in non-small cell lung cancer. , 2005, Cancer research.

[18]  G. V. Vande Woude,et al.  RNA Interference Reveals that Ligand-Independent Met Activity Is Required for Tumor Cell Signaling and Survival , 2004, Cancer Research.

[19]  J. Reis-Filho,et al.  Expression of c-met Tyrosine Kinase Receptor Is Biologically and Prognostically Relevant for Primary Cutaneous Malignant Melanomas , 2003, Oncology.

[20]  Kiyoko Kato,et al.  Hepatocyte growth factor modulates motility and invasiveness of ovarian carcinomas via ras mediated pathway , 2003, Molecular and Cellular Endocrinology.

[21]  L. Broemeling,et al.  Loss of MDA-7 expression with progression of melanoma. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[22]  K. Furge,et al.  Met receptor tyrosine kinase: enhanced signaling through adapter proteins , 2000, Oncogene.

[23]  I. Tsarfaty,et al.  Dominant negative Met reduces tumorigenicity-metastasis and increases tubule formation in mammary cells , 2000, Oncogene.

[24]  R. Sharp,et al.  c-Met autocrine activation induces development of malignant melanoma and acquisition of the metastatic phenotype. , 1998, Cancer research.

[25]  M. Fiscella,et al.  Evidence for a role of Met-HGF/SF during Ras-mediated tumorigenesis/metastasis , 1998, Oncogene.

[26]  M. Tsao,et al.  The roles of hepatocyte growth factor/scatter factor and met receptor in human cancers (Review). , 1998, Oncology reports.

[27]  P. Parsons,et al.  Detection of the c‐met proto‐oncogene product in normal skin and tumours of melanocytic origin , 1994, The Journal of pathology.

[28]  J. Hoeffler,et al.  Ras mutations in human melanoma: a marker of malignant progression. , 1994, The Journal of investigative dermatology.

[29]  F. McCormick,et al.  Structural requirements for the interaction of p21ras with GAP, exchange factors, and its biological effector target. , 1993, The Journal of biological chemistry.

[30]  C. Cooper,et al.  Mechanism of met oncogene activation , 1986, Cell.

[31]  R. Auerbach,et al.  Immunotherapy for Melanoma , 1973 .

[32]  L. Shun.  An Orally Available Small-Molecule Inhibitor of c-Met,PF-2341066,Exhibits Cytoreductive Antitumor Efficacy through Antiproliferative and Antiangiogenic Mechanisms , 2010 .

[33]  E. Grimm,et al.  NF-kappaB mediates mitogen-activated protein kinase pathway-dependent iNOS expression in human melanoma. , 2009, The Journal of investigative dermatology.

[34]  M. Sharpe,et al.  Hepatocyte growth factor/scatter factor (HGF/SF), the c-met receptor and the behaviour of epithelial cells. , 1993, Symposia of the Society for Experimental Biology.

[35]  R. Halaban,et al.  Met and hepatocyte growth factor/scatter factor signal transduction in normal melanocytes and melanoma cells. , 1992, Oncogene.