抗CD20嵌合抗体突变型Fab′片断诱导凋亡过程中活性氧Caspase-3的变化  

RAS and Caspase-3 Change in Apoptosis of B Lymphoma Cells Induced by One Amino Acid Mutation in An Anti-CD20 Antibody Fragment Fab′

作　　者：杨铭 范冬梅 刘银星 熊冬生 许元富 邵晓枫 杨纯正 

机构地区：[1]中国医学科学院协和医科大学血液学研究所血液病医院实验血液学国家重点实验室,天津300020

出　　处：《肿瘤防治研究》2005年第11期673-676,共4页Cancer Research on Prevention and Treatment

基　　金：国家863计划资助项目(2003AA215080);天津重大科技攻关资助项目(003119511)

摘　　要：目的研究抗CD20嵌合抗体突变型Fab′片断诱导Raji细胞凋亡过程中活性氧(ROS),Caspase3的变化。方法利用MTT法测定突变型Fab′片断抑制细胞生长,形态学方法观察凋亡细胞的变化,用流式细胞仪检测DCFHDA荧光探针标记细胞内ROS的变化,以及用酶标仪和Westernblot检测细胞内Caspase3的变化。结果MTT法测定突变型Fab′片断对Raji细胞的生长具有抑制作用,其抑制作用呈明显的剂量依赖性,荧光显微镜下观察细胞出现凋亡,流式细胞仪,酶标仪以及Westernblot检测细胞内ROS,Caspase3的表达增高,并与作用时间,剂量呈依赖关系。结论ROS,Caspase3表达的增高,在抗CD20嵌合抗体突变型Fab′片断诱导Raji细胞凋亡的过程中起到重要作用。：Objective To study effect of one amino acid mutation in an anti-cd20 antibody fragment Fab＇ on ROS, Caspase-3 in B lymphoma Raji cells. Methods MTT was used to observe the effect of anti-CD20 Fab＇ on Raji cell growth; morphologie change through microscope were used to assay apoptosiss induced by anti-CD20 Fab＇ ,ROS labeled with DCFH-DA in cells was processed on FACS, Caspase-3 gene expression local change in Raji cells FACS using Western-blottong. Results The result of MTT indicated that the growth of Raji cell was inhibited by chimeric anti-CD20 antibody fragment Fab＇, the apoptotie bodies were observed,ROS and Casepase-3 level in Raji cell was increased by anti-CD20 Fab＇. Conclusion The increasing of ROS,Caspase-3 level is important in the apoptosis of Raji cell induced by anti-CD20 Fab＇.

关 键 词：抗CD20抗体 随机突变 凋亡 ROS Caspase 

分 类 号：Q753[生物学—分子生物学]