HRAS mutations and resistance to the epidermal growth factor receptor tyrosine kinase inhibitor erlotinib in head and neck squamous cell carcinoma cells

Head Neck. 2014 Nov;36(11):1547-54. doi: 10.1002/hed.23499. Epub 2014 Mar 20.

Abstract

Background: The purpose of this study was to identify mechanisms of innate resistance to an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, erlotinib, in a panel of head and neck squamous cell carcinoma (HNSCC) cell lines. Specifically, we analyzed the role of HRAS mutations in erlotinib resistance.

Methods: Erlotinib sensitivity was determined by methyl thiazolyl-tetrazolium (MTT) assays. Molecular signaling pathways and somatic mutations were examined. Changes in sensitivity after modulation of HRAS expression were evaluated.

Results: All 7 cell lines were wild-type for EGFR and KRAS regardless of erlotinib sensitivity; however, 1 erlotinib-resistant cell line (HN31) harbored an HRAS G12D mutation. Downregulation of HRAS expression by small interfering RNA (siRNA) or short hairpin RNA (shRNA) in HN31 led to increased erlotinib sensitivity in vitro and in vivo. Transfection of activating HRAS-mutant (G12D and G12V) constructs into erlotinib-sensitive cell lines made them more resistant to erlotinib.

Conclusion: Activating HRAS mutations can confer erlotinib resistance in an HRAS mutant HNSCC cell line.

Keywords: HRAS; epidermal growth factor receptor; erlotinib; head and neck squamous cell carcinoma; resistance.

Publication types

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

MeSH terms

  • Animals
  • Blotting, Western
  • Carcinoma, Squamous Cell / drug therapy
  • Carcinoma, Squamous Cell / genetics
  • Cell Line, Tumor / drug effects
  • Cell Proliferation / drug effects
  • Down-Regulation / genetics
  • Drug Resistance, Neoplasm / genetics*
  • Erlotinib Hydrochloride
  • Head and Neck Neoplasms / drug therapy
  • Head and Neck Neoplasms / genetics
  • Humans
  • Mice
  • Molecular Targeted Therapy / methods*
  • Mutation*
  • Protein Kinase Inhibitors / pharmacology
  • Proto-Oncogene Proteins p21(ras) / drug effects
  • Proto-Oncogene Proteins p21(ras) / genetics*
  • Quinazolines / pharmacology*
  • Sensitivity and Specificity
  • Signal Transduction / drug effects
  • Squamous Cell Carcinoma of Head and Neck
  • Transfection

Substances

  • Protein Kinase Inhibitors
  • Quinazolines
  • Erlotinib Hydrochloride
  • HRAS protein, human
  • Proto-Oncogene Proteins p21(ras)