β-胡萝卜素辅致癌机制体外实验研究  

Co-carcinogenic Effect of β-Carotene on BALB/c-3T3 Cell Transformation Induced by Benzo(α)pyrene and Cigarette-smoke Condensate

作　　者：应贤平[1] 仲伟鉴[1] 肖萍[1] 郑志清[1] 董竞武[1] 郑勇英[1] 周祥凤[1] 

机构地区：[1]上海市疾病预防控制中心,上海200336

出　　处：《环境与职业医学》2003年第3期164-167,共4页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金资助 (编号 :39980 0 35)

摘　　要：[目的 ]建立改进的BALB/c 3T3细胞转化试验方法 ,简便、快速检测 β 胡萝卜素通过诱导细胞色素P45 0酶提高苯并 (a)芘、吸烟焦油致癌作用。 [方法 ]每组 12瓶细胞 ,每瓶接种 10 4细胞 ,2 4h后加受试物 ,1周后 2瓶细胞用结晶紫方法测定细胞毒性 ,其余 10瓶换含 2 %胎牛血清和 1%ITES( 2 0 0 μg/ml胰岛素 ,2 0 0 μg/ml人转铁蛋白 ,12 2 μg/ml氨基乙醇 ,0 0 3 4μg/ml亚硒酸钠 )DMEM /F12转化表达培养液。每周换液 2次 ,第 2 5天甲醇固定 10min ,2 5 %Giemsa染色 5～ 10min ,肉眼计数直径 >2mm转化灶。 [结果 ] 0 5、1μmol/Lβ 胡萝卜素、10 μmol/Lα 萘黄酮、10 μmol/L苯并 (a)芘和 2μg/ml吸烟焦油对细胞相对存活率均大于 90 %。 1μmol/Lβ 胡萝卜素能提高苯并 (a)芘、吸烟焦油的细胞转化率平均每瓶细胞转化灶分别增高 3 2 %和 5 0 0 %。α 萘黄酮可以抑制 β 胡萝卜素增强吸烟焦油和苯并 (a)芘细胞转化作用。[结论 ]β 胡萝卜素增强苯并 (a)芘和吸烟焦油对BALB/c 3T3细胞转化能力 ,可被混合功能氧化酶抑制剂α 萘黄酮阻止 ,提示 β 胡萝卜素辅致癌作用是通过诱导细胞色素P45Objective] To assess the β carotene property of inducing cytochrome P450 for bioactivating benzo(a)pyrene(B(a)P) and cigarette smoke condensate(TAR) using the improved BALB/c 3T3 cell transformation assay. [Methods] BALB/c 3T3 cells were plated at a density of 10 000 cells per 5 ml plate in twelve replicates per each treatment. After 24 hours,the cells were exposed to the test chemicals,and cultured for one week,then the cytotoxicity was determined. The remaning ten plates were incubated in fresh DMEM/F12 medium containing 2% serum and 1%ITES(200 μg/ml of insulin,200 μg/ml of transferrin,12 2 μg/ml of ethanolamine and 0 034 μg/ml of sodium selenite). The medium was changed twice a week. On 25 th day,the cells were fixed with methanol and stained with 2 5% Giemsa solution;>2 mm foci in diameter were counted. [Results] 0 1,1 μmol/L of β carotene,10 μmol/L of α naphthoflavone ,10 μmol/L of B(a)P and 2 μg/ml of TAR showed non toxic or only slightly toxic. Transformation frequency of B(a)P and TAR was markedly enhanced by 1 μmol/L of β carotene, and the formation of foci was significantly reduced in the presence of α naphthoflavone . [Conclusion] The enhanced effect of β carotene in the formation of transformation foci by B(a)P and TAR was inhibited by α naphthoflavone. This finding suggested that co carcinogenic effect of β carotene results from inducing cytochrome P450 enzyme.

关 键 词：Β-胡萝卜素 辅致癌作用 BALB/c-3T3细胞转化试验 检测 吸烟 抗癌 

分 类 号：R73－3[医药卫生—肿瘤]