Catechol estrogen conjugates and DNA adducts in the kidney of male Syrian golden hamsters treated with 4-hydroxyestradiol: potential biomarkers for estrogen-initiated cancer

• Published in: Carcinogenesis (New York) Vol. 22; no. 3; pp. 489 - 497
• Main Authors: Devanesan, Prabu, Todorovic, Rosa, Zhao, Jiang, Gross, Michael L., Rogan, Eleanor G., Cavalieri, Ercole L.
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.03.2001; Oxford Publishing Limited (England)
• Subjects: 17β-estradiol; 4-hydroxyestradiol; 4-quinone(s); Animals; Biological and medical sciences; Biomarkers, Tumor - metabolism; Carcinogenesis, carcinogens and anticarcinogens; catechol estrogen(s); catechol estrogen-3; CE-Q; Chemical agents; Chromatography, High Pressure Liquid; Cricetinae; Cys; cysteine; DNA Adducts - metabolism; Dose-Response Relationship, Drug; electrospray ionization; ESI; Estradiol - administration & dosage; Estradiol - analogs & derivatives; Estrogens, Catechol - metabolism; estrone; glutathione; glutathione moiety; GSH; high-performance liquid chromatography; HPLC; hydroxyestrogen(s); Kidney - drug effects; Kidney - metabolism; LC/MS; liquid chromatography/mass spectrometry; Male; Mass Spectrometry; Medical sciences; Mesocricetus; N-acetylcysteine; NAcCys; Neoplasms, Experimental - chemically induced; Neoplasms, Experimental - metabolism; OHE; SG; Tumors; Cysteine; Intraperitoneal administration; Toxicity; Estrogen; Acetylcysteine; Biological marker; Carcinogenesis; Kidney; Catechol estrogen; Molecular adduct; Glutathione; Kidney disease; Hormone; Urinary system disease; Rodentia; Tumor initiation; Malignant tumor; Metabolism; Carcinogen; Vertebrata; Mammalia; Animal; DNA; Lesion; Conjugated compound; Hamster
• ISSN: 0143-3334; 1460-2180; 1460-2180
• DOI: 10.1093/carcin/22.3.489

Formation of depurinating adducts by reaction of catechol estrogen-3,4-quinones with DNA was proposed to be a tumor initiating event by estrogens [E.L.Cavalieri et al. (1997) Proc. Natl Acad. Sci. USA, 94, 10937–10942]. Under estrogenic imbalance, oxidation of catechol estrogens to quinones may compete with their detoxification by protective enzymes. The quinones formed can be detoxified by reaction with glutathione (GSH) or can covalently bind to DNA. To provide more support for this hypothesis, we developed a method to identify and quantify GSH, cysteine (Cys) and N-acetylCys conjugates of 4-hydroxyestrogens (4-OHE) in the kidneys of male Syrian hamsters treated with 4-hydroxyestradiol (4-OHE2) by intraperitoneal injection. The highest level of conjugates was observed 1 h after treatment, and almost none was detected after 24 h. Dose–response studies indicated conjugate formation after treatment with 0.5 μmol of 4-OHE2/100 g body weight, and formation increased up to a treatment level of 12 μmol/100 g body weight. GSH, Cys and N-acetylCys conjugates of 4-OHE were identified in the picomole range by high-performance liquid chromatography (HPLC) with multichannel electrochemical detection and confirmed by HPLC/tandem mass spectrometry. Treatment of tissue homogenates with β-glucuronidase/sulfatase at 37°C for 6 h before extraction resulted in a 12- to 20-fold increase in Cys conjugates from picomole to nanomole levels. Similar enhancement was observed by just incubating the tissue at 37°C for 6 h. Evidence for the 4-OHE-1-N7Gua depurinating adducts was obtained by mass spectrometry. We conclude that GSH and Cys conjugates of the 4-OHE and the 4-OHE-N7Gua adducts can be utilized as biomarkers to detect estrogenic imbalance and potential susceptibility to tumor initiation.