title: Differential chemosensitivity to antifolate drugs between RAS and BRAF melanoma cells.
abstract: BACKGROUND
  The importance of the genetic background of cancer cells for the individual susceptibility to cancer treatments is increasingly apparent. In melanoma, the existence of a BRAF mutation is a main predictor for successful BRAF-targeted therapy. However, despite initial successes with these therapies, patients relapse within a year and have to move on to other therapies. Moreover, patients harbouring a wild type BRAF gene (including 25% with NRAS mutations) still require alternative treatment such as chemotherapy. Multiple genetic parameters have been associated with response to chemotherapy, but despite their high frequency in melanoma nothing is known about the impact of BRAF or NRAS mutations on the response to chemotherapeutic agents.
  
  
  METHODS
  Using cell proliferation and DNA methylation assays, FACS analysis and quantitative-RT-PCR we have characterised the response of a panel of NRAS and BRAF mutant melanoma cell lines to various chemotherapy drugs, amongst them dacarbazine (DTIC) and temozolomide (TMZ) and DNA synthesis inhibitors.
  
  
  RESULTS
  Although both, DTIC and TMZ act as alkylating agents through the same intermediate, NRAS and BRAF mutant cells responded differentially only to DTIC. Further analysis revealed that the growth-inhibitory effects mediated by DTIC were rather due to interference with nucleotide salvaging, and that NRAS mutant melanoma cells exhibit higher activity of the nucleotide synthesis enzymes IMPDH and TK1. Importantly, the enhanced ability of RAS mutant cells to use nucleotide salvaging resulted in resistance to DHFR inhibitors.
  
  
  CONCLUSIONS
  In summary, our data suggest that the genetic background in melanoma cells influences the response to inhibitors blocking de novo DNA synthesis, and that defining the RAS mutation status could be used to stratify patients for the use of antifolate drugs.
authors.affiliation: Manchester Cancer Research Centre, The University of Manchester, Michael Smith Building, Oxford Road, Manchester M13 9PT, UK. Imanol.Arozarena@manchester.ac.uk.
authors.forename: Imanol
authors.initials: I
authors.lastname: Arozarena
authors.forename: Ibai
authors.initials: I
authors.lastname: Goicoechea
authors.forename: Oihane
authors.initials: O
authors.lastname: Erice
authors.forename: Jennnifer
authors.initials: J
authors.lastname: Ferguson
authors.forename: Geoffrey P
authors.initials: GP
authors.lastname: Margison
authors.forename: Claudia
authors.initials: C
authors.lastname: Wellbrock
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D019175:DNA Methylation / Q000235:genetics; D003606:Dacarbazine / Q000008:administration & dosage / Q000031:analogs & derivatives; D004791:Enzyme Inhibitors / Q000008:administration & dosage; D020558:GTP Phosphohydrolases / Q000235:genetics*; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D008565:Membrane Proteins / Q000235:genetics*; D009154:Mutation; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D012878:Skin Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D000077204:Temozolomide
chemical_list: D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D003606:Dacarbazine; D004247:DNA; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D020558:GTP Phosphohydrolases; C579846:NRAS protein, human; D000077204:Temozolomide