Somatic mutation analysis in melanoma using targeted next generation sequencing

Exp Mol Pathol. 2017 Oct;103(2):172-177. doi: 10.1016/j.yexmp.2017.08.006. Epub 2017 Aug 16.


Advanced stage malignant melanoma often responds poorly to therapy with low survival rates. New therapeutic approaches are based upon a growing understanding of the underlying molecular abnormalities. We demonstrate the feasibility of a next generation sequencing (NGS) assay, which targets hotspots in 50 cancer genes, to assess genotypes that may influence therapeutic selection and response. DNA was extracted from formalin fixed paraffin embedded (FFPE) melanoma specimens to create multiplexed libraries which were sequenced. Of the 121 cases, BRAF mutations were present in 48 cases (40%) and NRAS mutations in 24 cases (20%). We identified other gene variants in 20 BRAF-mutated cases. Additional gene variants were also identified in the 57 BRAF wild-type cases. Four patients harbored different gene mutations at metastatic sites as compared to their primary lesions or metastasis from different sites. Concurrent gene variants may provide additional targets for future therapies and may suggest alternative mechanisms of secondary resistance.

Keywords: BRA; Melanoma; Mutation; NRAS; Next generation sequencing (NGS); Variant.

MeSH terms

  • Biomarkers, Tumor / genetics*
  • DNA Mutational Analysis / methods*
  • Female
  • GTP Phosphohydrolases / genetics*
  • Genotype
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Male
  • Melanoma / diagnosis*
  • Melanoma / genetics
  • Membrane Proteins / genetics*
  • Middle Aged
  • Mutation*
  • Neoplasm Metastasis
  • Paraffin Embedding
  • Prognosis
  • Proto-Oncogene Proteins B-raf / genetics*
  • Retrospective Studies


  • Biomarkers, Tumor
  • Membrane Proteins
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • GTP Phosphohydrolases
  • NRAS protein, human