Single CpG site methylation controls estrogen receptor gene transcription and correlates with hormone therapy resistance

J Steroid Biochem Mol Biol. 2017 Jul:171:209-217. doi: 10.1016/j.jsbmb.2017.04.001. Epub 2017 Apr 12.

Abstract

Hormone therapy is the most effective treatment for patients with estrogen receptor α-positive breast cancers. However, although resistance occurs during treatment in some cases and often reflects changed estrogen receptor α status, the relationship between changes in estrogen receptor α expression and resistance to therapy are poorly understood. In this study, we identified a mechanism for altered estrogen receptor α expression during disease progression and acquired hormone therapy resistance in aromatase inhibitor-resistant breast cancer cell lines. Subsequently, we investigated promoter switching and DNA methylation status of the estrogen receptor α promoter, and found marked changes of methylation at a single CpG site (CpG4) in resistant cells. In addition, luciferase reporter assays showed reduced transcriptional activity from this methylated CpG site. This CpG region was also completely conserved among species, suggesting that it acts as a methylation-sensitive Ets-2 transcription factor binding site, as confirmed using chromatin immunoprecipitation assays. In estrogen receptor α-positive tumors, CpG4 methylation levels were inversely correlated with estrogen receptor α expression status, suggesting that single CpG site plays an important role in the regulation of estrogen receptor α transcription.

Keywords: Breast cancer; DNA methylation; Estrogen receptor alpha; Ets-2; Hormone therapy resistance.

MeSH terms

  • Antineoplastic Agents, Hormonal / pharmacology
  • Aromatase Inhibitors / pharmacology
  • Base Sequence
  • Breast Neoplasms / drug therapy
  • Breast Neoplasms / metabolism*
  • Conserved Sequence
  • DNA Methylation* / drug effects
  • DNA, Recombinant / metabolism
  • Dinucleoside Phosphates / metabolism*
  • Drug Resistance, Neoplasm
  • Estrogen Receptor alpha / chemistry
  • Estrogen Receptor alpha / genetics
  • Estrogen Receptor alpha / metabolism*
  • Female
  • Genes, Reporter / drug effects
  • Humans
  • MCF-7 Cells
  • Middle Aged
  • Mutation
  • Neoplasm Proteins / chemistry
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Promoter Regions, Genetic* / drug effects
  • Proto-Oncogene Protein c-ets-2 / genetics
  • Proto-Oncogene Protein c-ets-2 / metabolism*
  • Recombinant Proteins / metabolism
  • Response Elements / drug effects
  • Transcription, Genetic* / drug effects

Substances

  • Antineoplastic Agents, Hormonal
  • Aromatase Inhibitors
  • DNA, Recombinant
  • Dinucleoside Phosphates
  • ESR1 protein, human
  • ETS2 protein, human
  • Estrogen Receptor alpha
  • Neoplasm Proteins
  • Proto-Oncogene Protein c-ets-2
  • Recombinant Proteins
  • cytidylyl-3'-5'-guanosine