活化蛋白-1在硒拮抗砷致细胞周期改变中的作用  

The Antagonism of Selenium on Arsenic-induced Cell Cycle Changes by Activator Protein-1 Regulation

作　　者：王艺陪[1,2] 李晓暖[1] 来瑞平[1] 余日安[3] 鲁文清[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院劳动卫生与环境卫生学系教育部环境与健康重点实验室,湖北武汉430030 [2]上海大学环境与化学工程学院污染与健康研究所,上海200444 [3]广东药学院公共卫生学院劳动卫生与环境卫生学系,广东广州510310

出　　处：《癌变．畸变．突变》2009年第3期210-213,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金资助项目(30471500)

摘　　要：背景与目的:研究激活蛋白1(activator protein-1,AP-1)在硒拮抗砷致细胞周期改变中的作用。材料与方法:以小鼠皮肤JB6-CI41细胞为靶细胞,分别用亚硒酸钠(2.5μmol/L)和不同浓度的亚砷酸(3.125、6.25和12.5μmol/L)单独及联合染毒JB6-CI41细胞,并用AP-1特异化学抑制剂姜黄素(20μmol/L)与亚砷酸(12.5μmol/L)联合染毒细胞,实验同时设生理盐水溶剂对照。上述各组染毒2 h后收获细胞,用凝胶阻滞电泳技术(electrophoretic mobility shift assay,EMSA)检测细胞AP-1的DNA结合活性,用流式细胞仪检测细胞周期的变化。观察砷诱导细胞周期改变与AP-1的DNA结合活性之间的关系。结果:与溶剂对照相比,各浓度的亚砷酸均可引起JB6-CI41细胞AP-1的DNA结合活性显著增加(P<0.01)和G1期比率显著性减少(P<0.01),较高浓度的亚砷酸(6.25和12.5μmol/L)引起G2期比率增多(P<0.05,P<0.01),2.5μmol/L亚硒酸钠对AP-1的DNA结合活性和细胞周期均无显著影响(P均>0.05)。亚硒酸钠与亚砷酸联合作用时,与对应浓度的亚砷酸单独作用相比,JB6-CI41细胞的AP-1的DNA结合活性明显降低(P<0.01),G1比率明显增多(P<0.01),G2期比率显著减少(P<0.05)。AP-1抑制剂姜黄素与亚砷酸联合染毒与对应的亚砷酸单独染毒组相比,AP-1的DNA结合活性明显降低(P<0.01),G1期比率增高(P<0.01)而G2期比率降低(P<0.01)。结论:砷通过激活AP-1通路影响细胞周期的变化;硒在一定浓度下能够通过拮抗砷对AP-1的活化而抑制砷引起的细胞周期变化,这可能是硒拮抗砷致癌作用的机制之一。BACKGROUND AND AIM： To explore the effect of selenium on arsenic-induced cell cycle changes by regulation of activator protein-1 （AP-1） MATERIALS AND METHODS： JB6-CI41 cells were exposed to sodium selenite alone at the concentrations of 2.5 μmol/L or combined with arsenous acid at the concentrations of 3.125, 6.25, and 12.5 μmol/ L. The AP-1 DNA binding activity and cell cycle were analyzed by electrophoretic mobility shift assay （EMSA） and flow cytometry. Meanwhile, curcumin （20 μmol/L）, a specific inhibitor of AP-1, was used to treat JB6 cells together with arsenous acid（ 12.5 μmol/L） in order to understand the relationship between arsenic-induced cell cycle changes and AP-1 DNA binding activity. RESULTS： Compared with solvent control, arsenous acid （3. 125, 6.25 and 12.5μmol/ L） could significantly up-regulate AP-1 DNA binding activity and reduce G1 phase cells （P〈0.01, P 〈 0.01）, and higher concentrations of arsenous acid （6.25 and 12.5 μmol/L） increased G2 phase cells （P 〈0.05, P〈0.01） . Sodium selenite （2.5μmol/L） showed no effect on AP-1 DNA binding activity and cell cycle（P〉0.05, P〉 0.05） Compared with corresponding concentrations of arsenous acid, co-exposure of JB6-CI41 cells to sodium selenite （2.5 μmol/L） and arsenous acid （3.125,6.25 and 12.5 μmol/L） down-regulated AP-1 DNA binding activity（P 〈 0.01） , increased G1 phase cells （P 〈 0.01）, and decreased G2 phase cells （P 〈 0.05） . Co-exposure of JB6-CI41 cells to curcumin and arsenous acid could significantly down-regulate AP-1 DNA binding activity, increased G, phase cells and decreased G2 phase cells compared with corresponding concentrations of arsenous acid（all P〈0.01） CONCLUSION： Arsenic may change ceil cycle by activating AP-1 signal pathway. Certain concentration of selenium could down-regulate arsenic-induced AP-1 DNA binding activity, which may be one of the mechanisms that selenium could antagonize the carcinogenetic property of arsenic.

关 键 词：砷 硒 AP-1 细胞周期 致癌作用 

分 类 号：R135.1[医药卫生—劳动卫生]