Expression Profiling of a Human Thyroid Cell Line Stably Expressing the BRAFV600E Mutation

Cancer Genomics Proteomics. 2017 Jan 2;14(1):53-67. doi: 10.21873/cgp.20018.

Abstract

Background/aim: The BRAFV600E mutation acts as an initiator of cancer development in papillary thyroid carcinoma (PTC). Gene expression changes caused by the BRAFV600E mutation may have an important role in thyroid cancer development.

Materials and methods: To study genomic alterations caused by the BRAFV600E mutation, we made human thyroid cell lines that harbor the wild-type BRAF gene (Nthy/WT) and the V600E mutant-type BRAF gene (Nthy/V600E).

Results: Flow cytometry and western blotting showed stable transfection of the BRAF gene. In functional experiments, Nthy/V600E showed increased anchorage-independent growth and invasion through Matrigel, compared to Nthy/WT. Microarray analysis revealed that 2,441 genes were up-regulated in Nthy/V600E compared to Nthy/WT. Gene ontology analysis showed that the up-regulated genes were associated with cell adhesion, migration, and the ERK and MAPK cascade, and pathway analysis showed enrichment in cancer-related pathways.

Conclusion: Our Nthy/WT and Nthy/V600E cell line pair could be a suitable model to study the molecular characteristics of BRAFV600E PTC.

Keywords: BRAFV600E; Nthy-ori 3-1 cell; Thyroid cancer; microarrays; signal transduction.

MeSH terms

  • Carcinoma / genetics
  • Carcinoma / metabolism
  • Carcinoma, Papillary
  • Cell Line
  • Cell Movement / genetics
  • Codon*
  • Female
  • Flow Cytometry
  • Gene Expression Profiling*
  • Gene Expression Regulation
  • Gene Expression*
  • Humans
  • Male
  • Mutation*
  • Proto-Oncogene Proteins B-raf / genetics*
  • Proto-Oncogene Proteins B-raf / metabolism
  • Sequence Analysis, DNA
  • Thyroid Cancer, Papillary
  • Thyroid Gland / metabolism*
  • Thyroid Neoplasms / genetics
  • Thyroid Neoplasms / metabolism
  • Transduction, Genetic

Substances

  • Codon
  • Proto-Oncogene Proteins B-raf