Mutagenic properties of estrogen quinone-derived DNA adducts in simian kidney cells.
Mutagenic properties of estrogen quinone-derived DNA adducts in simian kidney cells.
复制标题
猿猴肾细胞中雌激素醌衍生的 DNA 加合物的诱变特性。
DOI：
10.1021/bi002273c
复制
发表时间：
2001
期刊：
影响因子：
2.9
通讯作者：
Shibutani,S
中科院分区：
文献类型：
--
作者：
Terashima,I;Suzuki,N;Shibutani,S
儿茶酚雌激素引起的DNA损伤已被证明在肿瘤形成中起病因作用。儿茶酚雌激素与DNA反应，并通过醌形式形成几种DNA加合物。为了探索2-羟基雌激素衍生的DNA加合物在哺乳动物细胞中的致突变特性，N2-（2-羟基雌激素-6-基）-2 '-脱氧鸟苷和N6-由2-羟基雌酮、2-羟基雌酮或2-羟基雌三醇的醌类诱导的（2-羟基雌激素-6-基）-2 '-脱氧腺苷加合物被位点特异性地掺入到寡脱氧核苷酸中。（5 ′ TCCTCCTCXCCTCTC，其中X是dG、dA、2-OHE-N2-dG或2-OHE-N6-dA）。将修饰的寡核苷酸插入单链噬菌粒载体中，然后转染到猴肾（COS-7）细胞中。相对于所有2-OHE-N2-dG加合物，观察到dCMP（正确的碱基）的优先掺入。仅检测到靶向G → T颠换;在2-OHE 2-N2-dG加合物对面观察到最高突变频率（18.2%），其次是2-OHE 1-N2-dG（4.4%）和2-OHE 3-N2-dG（1.3%）。当使用2-OHE-N6-dA加合物时，观察到dTMP（正确的碱基）的优先掺入。检测到代表A → T颠换的靶向突变，伴有少量A → G转换。2-OHE 1-N6-dA和2-OHE 3-N6-dA的突变频率最高（分别为14.5%和14.1%），而2-OHE 2-N6-dA的突变频率仅为6.0%。用含有未修饰的寡脱氧核苷酸的载体未检测到突变。因此，2-OHE醌衍生的DNA加合物具有致突变性，主要在哺乳动物细胞中产生G → T和A → T突变。突变频率根据2-OHE部分的性质而变化。
DNA damage caused by catechol estrogens has been shown to play an etiologic role in tumor formation. Catechol estrogens are reactive to DNA and form several DNA adducts via their quinone forms. To explore the mutagenic properties of 2-hydroxyestrogen-derived DNA adducts in mammalian cells,N2-(2-hydroxyestrogen-6-yl)-2‘-deoxyguanosine andN6-(2-hydroxyestrogen-6-yl)-2‘-deoxyadenosine adducts induced by quinones of 2-hydroxyestrone, 2-hydroxyestradiol, or 2-hydroxyestriol were incorporated site-specifically into the oligodeoxynucleotides (5‘TCCTCCTCXCCTCTC, where X is dG, dA, 2-OHE-N2-dG, or 2-OHE-N6-dA). The modified oligodeoxynucleotides were inserted into single-stranded phagemid vectors followed by transfection into simian kidney (COS-7) cells. Preferential incorporation of dCMP, the correct base, was observed opposite all 2-OHE-N2-dG adducts. Only targeted G → T transversions were detected; the highest mutation frequency (18.2%) was observed opposite the 2-OHE2-N2-dG adduct, followed by 2-OHE1-N2-dG (4.4%) and 2-OHE3-N2-dG (1.3%). When 2-OHE-N6-dA adducts were used, preferential incorporation of dTMP, the correct base, was observed. Targeted mutations representing A → T transversions were detected, accompanied by small numbers of A → G transitions. The highest mutation frequencies were observed with 2-OHE1-N6-dA and 2-OHE3-N6-dA (14.5 and 14.1%, respectively), while 2-OHE2-N6-dA exhibited a mutation frequency of only 6.0%. No mutations were detected with vectors containing unmodified oligodeoxynucleotides. Thus, 2-OHE quinone-derived DNA adducts are mutagenic, generating primarily G → T and A → T mutations in mammalian cells. The mutational frequency varied depending on the nature of the 2-OHE moiety.