Characterization of estrogen quinone-derived protein adducts and their identification in human serum albumin derived from breast cancer patients and healthy controls |
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| Authors: | Chen Dar-Ren Chen Shou-Tung Wang Tzu-Wen Tsai Chen-His Wei Hz-Han Chen Guan-Jie Yang Tsung-Chou Lin Che Lin Po-Hsiung |
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| Affiliation: | a Comprehensive Breast Cancer Center, Changhua Christian Hospital, Changhua, Taiwan
b Department of Environmental Engineering, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 402, Taiwan |
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| Abstract: | Both 17β-estradiol-2,3-quinone (E2-2,3-Q) and 17β-estradiol-3,4-quinone (E2-3,4-Q) are reactive metabolites of estrogen that are thought to be responsible for the estrogen-induced genotoxicity. The aim of this study was to establish a methodology to analyze estrogen quinone-derived protein adducts and to measure the background levels of these adducts in human serum albumin (Alb) derived from female blood donors in Taiwan. Results from in vitro experiments confirmed that the production of estrogen quinone-derived adducts on serum Alb increased with increased concentration of estrogen quinones. Time-course experiments suggested that both E2-2,3-Q- and E2-3,4-Q-derived adducts rapidly reached maximum values at 10 min mark and remained constant thereafter for up to 24 h. Additionally, with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) pretreatment, the production of estrogen quinone-derived protein adducts was detected in human MCF-7 breast cancer cells exposed to estrogen. Co-treatment of a catechol-O-methyl transferase inhibitor further enhanced the production of estrogen quinone-derived adducts in all cases. When we investigated the levels of estrogen quinone-derived adducts in human serum Alb, cysteinyl adducts of E2-2,3-Q-1-S-Alb, E2-2,3-Q-4-S-Alb, and E2-3,4-Q-2-S-Alb were detected in all healthy female controls (n = 10) with median levels at 147 (range 14.1-533), 197 (range 30.0-777), and 65.6 (range 17.6-1360) (pmol/g), respectively. We noticed that levels of E2-2,3-Q-derived adducts were 2-fold greater than those of E2-3,4-Q-2-S-Alb in controls whereas levels of E2-3,4-Q-2-S-Alb were 2-fold higher than those of E2-2,3-Q-derived adducts in patients (n = 20). Additionally, levels of E2-2,3-Q-4-S-Alb correlated significantly with those of E2-3,4-Q-2-S-Alb (correlation coefficient r = 0.684-0.850, p < 0.05). Overall, we conclude that cumulative body burden of E2-3,4-Q is a significant predictor of breast cancer. |
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| Keywords: | AhR, arylhydrocarbon receptor Alb, albumin COMT, catechol-O-methyl transferase Cp, cytoplasmic proteins CYP1A1, cytochrome P450 1A1 CYP1B1, cytochrome P450 1B1 DMSO, dimethyl sulfoxide E2, 17β-estradiol E2-2,3-Q, 17β-estradiol-2,3-quinone E2-3,4-Q, 1,7β-estradiol-3,4-quinone E2-2,3-Q-1-NAC and E2-2,3-Q-4-NAC, reaction products of E2-2,3-Q with N-acetyl- smallcaps" >l-cysteine E2-3,4-Q-2-NAC, reaction products of E2-3,4-Q with N-acetyl- smallcaps" >l-cysteine E2-2,3-Q-1-S-Alb and E2-2,3-Q-1-S-Alb, adducts resulting from reaction of E2-2,3-Q with Alb E2-3,4-Q-2-S-Alb, adducts resulting from reaction of E2-3,4-Q with Alb E2-2,3-Q-1-S-Hb and E2-2,3-Q-4-S-Hb, adducts resulting from reaction of E2-2,3-Q with Hb E2-3,4-Q-2-S-Hb, adducts resulting from reaction of E2-3,4-Q with Hb E2-2,3-Q-1-S-TFA and E2-2,3-Q-4-S-TFA, trifluoroacetyl derivative of E2-2,3-Q adduct after adduct cleavage E2-3,4-Q-2-S-TFA, trifluoroacetyl derivatives of E2-3,4-Q adduct after adduct cleavage EI, electron impact ERα, estrogen receptors-alpha GC-MS, gas chromatograph and mass spectrometer [2H5]-E2, E2-2, 4, 16, 16, 17-d5 ([2H5]-E2) Hb, hemoglobin HPLC, high performance liquid chromatography MSA, methanesulfonic acid MT assay, methanesulfonic acid and trifluoroacetic acid anhydride assay NAC, N-acetyl- smallcaps" >l-cysteine NICI, negative ion chemical ionization ROS, reactive oxygen species SD, standard deviation TCDD, 2,3,7,8-tetrachlorodibenzo-p-dioxin TFA, trifluoroacetyl TFAA, trifluoroacetic acid anhydride |
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