Differential responses of human papillary thyroid cancer cell lines carrying the RET/PTC1 rearrangement or a BRAF mutation to MEK1/2 inhibitors

Arch Otolaryngol Head Neck Surg. 2007 Aug;133(8):810-5. doi: 10.1001/archotol.133.8.810.

Abstract

Objectives: To examine the effects of 2 mitogen-activated protein kinase kinase (MEK1/2) inhibitors on papillary thyroid carcinoma (PTC) cell lines carrying the RET/PTC1 rearrangement or a BRAF mutation. In PTC, RET/PTC1 rearrangement or BRAF mutations results in constitutional activation of RET kinase or BRAF, respectively. Along the RET or BRAF signaling cascades, the activated RET kinase or BRAF activates MEK1/2, and then mitogen-activated protein kinases (extracellular signal-related kinase 1/2 [ERK1/2]) is activated. Activated ERK1/2 enters the nucleus and phosphorylates a variety of transcription factors, resulting in cancer cell proliferation. The MEK1/2 inhibitors, PD98059 and U0126, have been shown to inhibit cell growth in other cancers.

Design: In vitro study.

Subjects: Papillary thyroid carcinoma cell lines carrying the RET/PTC1 rearrangement (BHP2-7) or a BRAF mutation (BHP5-16).

Intervention: We treated PTC cells carrying the RET/PTC1 rearrangement or a BRAF mutation with 2 MEK1/2 inhibitors (PD98059 and U0126).

Main outcome measures: Using Western blot analysis, we detected the expression of phosphorylated ERK1/2 and expression of cleaved poly(ADP-ribose) polymerase (PARP) in cells after treatment with either inhibitors. Growth inhibition was monitored by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

Results: Using Western blot analysis, we detected the dephosphorylation of ERK1/2 in PTC cells carrying the RET/PTC1 rearrangement or a BRAF mutation after treating the cells with 2 MEK1/2 inhibitors (PD98059 and U0126). In addition, both PD98059 and U0126 completely inhibited the growth of the PTC cells carrying a BRAF mutation but partially inhibited the growth of the PTC cells carrying the RET/PTC1 rearrangement. Finally, we observed PARP cleavage only in cells with a BRAF mutation in the Western blot analysis.

Conclusion: These data suggested that treatment with MEK1/2 inhibitors can be used as tools for inhibiting the growth of PTC cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Butadienes / pharmacology
  • Butadienes / therapeutic use*
  • Carcinoma, Papillary / drug therapy*
  • Carcinoma, Papillary / genetics*
  • Cell Division / drug effects
  • Cell Line, Tumor
  • Flavonoids / pharmacology
  • Flavonoids / therapeutic use*
  • Gene Rearrangement
  • Humans
  • Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors*
  • Nitriles / pharmacology
  • Nitriles / therapeutic use*
  • Phosphorylation
  • Point Mutation
  • Protein Kinase Inhibitors / pharmacology
  • Protein Kinase Inhibitors / therapeutic use*
  • Proto-Oncogene Proteins B-raf / genetics*
  • Proto-Oncogene Proteins c-ret / genetics*
  • Thyroid Neoplasms / drug therapy*
  • Thyroid Neoplasms / genetics*

Substances

  • Butadienes
  • Flavonoids
  • Nitriles
  • Protein Kinase Inhibitors
  • U 0126
  • Proto-Oncogene Proteins c-ret
  • RET protein, human
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • Mitogen-Activated Protein Kinase 1
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one