Formation of depurinating N3Adenine and N7Guanine adducts by MCF-10F cells cultured in the presence of 4-hydroxyestradiol

Muhammad Saeed, Eleanor G Rogan, Sandra V. Fernandez, Fathima Sheriff, Jose Russo, Ercole Cavalieri

Research output: Contribution to journalArticle

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Abstract

Metabolic conversion of endogenous estrogens, estradiol (E2) and estrone (E1), to the catechol estrogens 4-hydroxyE 1(E2) [4-OHE1(E2)] has been implicated in the initiation of cancer in rodents and humans. Evidence collected in our laboratories has shown that 4-OHE1(E2) are enzymatically oxidized to E1(E2)-3,4-quinones [E 1(E2)-3,4-Q], which have the potential to damage DNA by forming predominantly depurinating adducts, 4-OHE1(E 2)-1-N3Ade and 4-OHE1(E2)-1-N7Gua, leading to the accumulation of mutations and probably cell transformation. The human breast epithelial cell line MCF-10F has been transformed by treatment with E 2 or 4-OHE2. We have used MCF-10F cells to study the presence of adducts and conjugates after treatment with 4-OHE2. To mimic the intermittent exposure of breast cells to endogenous estrogens, MCF-10F cells were treated with 1 μM 4-OHE2 for a 24-h period at 72, 120, 192 and 240 h postplating. Culture media were collected at each point, extracted by solid-phase extraction and analyzed by HPLC connected with a multichannel electrochemical detector and/or ultraperformance liquid chromatography/tandem mass spectrometry. Media from successive treatments with 4-OHE2 showed the formation of methoxy and cysteine conjugates, and the depurinating adducts 4-OHE1(E2)-1-N3Ade. The amount of 4-OHE1(E2)-1-N3Ade adducts was higher after the third treatment; smaller amounts of the 4-OHE1(E2)-1-N7Gua adducts were detected after the second and third treatments. These results demonstrate that MCF-10F cells oxidize 4-OHE2 to E1(E 2)-3,4-Q, which react with DNA to form the depurinating N3Ade and N7Gua adducts. This DNA damage can play an important role in the 4-OHE 2-induced mutations and transformation of MCF-10F cells to malignant cells.

Original languageEnglish (US)
Pages (from-to)1821-1824
Number of pages4
JournalInternational Journal of Cancer
Volume120
Issue number8
DOIs
StatePublished - Apr 15 2007

Fingerprint

Cultured Cells
DNA Damage
Estrogens
Breast
Catechol Estrogens
Quinones
Estrone
Solid Phase Extraction
Tandem Mass Spectrometry
4-hydroxyestradiol
Liquid Chromatography
Cysteine
Culture Media
Estradiol
Rodentia
Epithelial Cells
High Pressure Liquid Chromatography
Cell Line
Mutation
DNA

Keywords

  • Breast cancer initiation
  • Catechol estrogen conjugates
  • Catechol estrogen quinines
  • Depurinating estrogen-DNA adducts
  • Estrogen metabolism
  • Human breast epithelial cells

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Formation of depurinating N3Adenine and N7Guanine adducts by MCF-10F cells cultured in the presence of 4-hydroxyestradiol. / Saeed, Muhammad; Rogan, Eleanor G; Fernandez, Sandra V.; Sheriff, Fathima; Russo, Jose; Cavalieri, Ercole.

In: International Journal of Cancer, Vol. 120, No. 8, 15.04.2007, p. 1821-1824.

Research output: Contribution to journalArticle

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