活性氧诱发人类11号染色体基因突变  

Mutagenicities of Human Chromosome 11 in A_L Cells Induced by Reactive Oxygen Species

作　　者：朱茂祥[1] 杨陟华[1] 龚诒芬[1] 曹珍山[1] 陆颖[1] Tom K.HEI 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]美国哥伦亚大学放射生物研究中心,ny10032

出　　处：《中国生物化学与分子生物学报》2002年第1期92-98,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助 (No .3970 0 0 30 )

摘　　要：对体外产生的和内源性刺激产生的活性氧 (ROS)诱发人类 11号染色体 (Hchr 11)基因突变规律及其突变谱进行研究 .体外羟自由基 (·OH)用过氧化氢 (H2 O2 )与Fe2 + 反应产生 ,并用化学发光(CL)进行相对定量分析 ;内源性ROS用佛波醇酯 (PMA)刺激人外周血白细胞产生 ,并用CL和特异性抗氧化物检测和鉴定 ;用包含单条Hchr 11的人 中国仓鼠卵巢细胞 (AL)为靶 ,经CD59表面抗原抗体筛选突变细胞克隆 ,研究ROS诱发的Hchr 11基因突变 ;突变克隆细胞DNA用Hchr 11上 5种标志基因引物进行多重PCR分析 ,结合琼脂糖凝胶电泳绘制基因突变谱 .结果表明 ,体外ROS可诱发Hchr 11基因突变 ,且·OH诱发基因突变的能力明显强于H2 O2 ,两者的突变谱也存在明显差异 ;PMA可刺激人外周血白细胞产生大量的多种ROS ,并诱发Hchr 11基因突变 ,突变谱综合了H2 O2 和·OH的所有特征 ;一些抗氧化物对内源性产生的ROS诱发Hchr 11基因突变有明显抑制作用 .提示体外和内源性ROS可诱发Hchr 11基因突变 。Mutagenicities and spectra of mutants of human chromosome 11 in the human hamster hybrid (A L cells) induced by exogenetic and endogenetic reactive oxygen species (ROS) were investigated. Hydroxyl radicals (·OH) were generated by hydrogen peroxides (H 2O 2) reacting with ferrous chloride in vitro, and quantitatively determinated by chemiluminescence (CL). Endogenetic ROS were produced by phorbol myristate acetate (PMA) stimulating human polymorphonuclear leukocytes (PMN), detected by CL and identified with specific antioxidants. A L cells were used for assaying ROS induced mutagenicities of Hchr 11 through screening mutants in presence of anti CD 59 surface antigen antibody (S1) and complement. Spectra of mutants were drawn through multiplex polymerase chain reaction (PCR) of DNA extracted from S1 mutants. Results showed that mutagenicities of Hchr 11 induced by ·OH were greater than H 2O 2, and their spectra of mutants were obviously different. PMA could stimulate PMN to produce a great deal of ROS, and caused mutagenicities of Hchr 11, and its spectra of mutants showed both features of spectra of mutants induced by ·OH and H 2O 2. Some antioxidants such as superoxide dismutase (SOD), catalase and mannitol had remarkably inhibition to ROS induced mutagenicities. It is indicated that exogenetic and endogenetic ROS could cause mutagenisities of Hchr 11, and various ROS was likely to induce different types of gene mutation.

关 键 词：活性氧 人类11号染色体 基因突变 突变谱 癌变 

分 类 号：R730.2[医药卫生—肿瘤] R394.6[医药卫生—临床医学]