MET inhibition in tumor cells by PHA665752 impairs homologous recombination repair of DNA double strand breaks

Int J Cancer. 2012 Feb 1;130(3):728-34. doi: 10.1002/ijc.26058. Epub 2011 May 9.

Abstract

Abnormal activation of cellular DNA repair pathways by deregulated signaling of receptor tyrosine kinase systems has broad implications for both cancer biology and treatment. Recent studies suggest a potential link between DNA repair and aberrant activation of the hepatocyte growth factor receptor Mesenchymal-Epithelial Transition (MET), an oncogene that is overexpressed in numerous types of human tumors and considered a prime target in clinical oncology. Using the homologous recombination (HR) direct-repeat direct-repeat green fluorescent protein ((DR)-GFP) system, we show that MET inhibition in tumor cells with deregulated MET activity by the small molecule PHA665752 significantly impairs in a dose-dependent manner HR. Using cells that express MET-mutated variants that respond differentially to PHA665752, we confirm that the observed HR inhibition is indeed MET-dependent. Furthermore, our data also suggest that decline in HR-dependent DNA repair activity is not a secondary effect due to cell cycle alterations caused by PHA665752. Mechanistically, we show that MET inhibition affects the formation of the RAD51-BRCA2 complex, which is crucial for error-free HR repair of double strand DNA lesions, presumably via downregulation and impaired translocation of RAD51 into the nucleus. Taken together, these findings assist to further support the role of MET in the cellular DNA damage response and highlight the potential future benefit of MET inhibitors for the sensitization of tumor cells to DNA damaging agents.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • BRCA2 Protein / metabolism
  • Cell Cycle / drug effects
  • Cell Line, Transformed
  • Cell Line, Tumor
  • DNA Breaks, Double-Stranded / drug effects*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Indoles / pharmacology*
  • Mice
  • Mutation
  • NIH 3T3 Cells
  • Neoplasms / genetics
  • Neoplasms / metabolism
  • Phosphorylation / drug effects
  • Proto-Oncogene Proteins c-met / antagonists & inhibitors*
  • Proto-Oncogene Proteins c-met / genetics
  • Rad51 Recombinase / metabolism
  • Recombinational DNA Repair / drug effects*
  • Sulfones / pharmacology*

Substances

  • 5-((2,6-dichlorobenzyl)sulfonyl)-3-((3,5-dimethyl-4-((2-(pyrrolidin-1-ylmethyl)pyrrolidin-1-yl)carbonyl)-1H-pyrrol-2-yl)methylene)-1,3-dihydro-2H-indol-2-one
  • Antineoplastic Agents
  • BRCA2 Protein
  • Indoles
  • Sulfones
  • MET protein, human
  • Proto-Oncogene Proteins c-met
  • Rad51 Recombinase