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

doi:10.3971/j.issn.1000-8578.3111

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 BLymphoma Cells Induced by One Amino Acid Mutation in An Anti-CD20 Antibody Fragment Fab′

摘要

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），Caspase-3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DCFH—DA荧光探针标记细胞内I的S的变化，以及用酶标仪和Westernblot检测细胞内Caspase-3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，Caspase-3的表达增高，并与作用时间，剂量呈依赖关系。结论 ROS，Caspase-3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。

Abstract: Objective 　To study effect of one amino acid mutation in an anti-cd20 antibody fragment Fab′on ROS, Caspase-3 in Blymphoma Raji cells. Methods 　MTT was used to observe the effect of anti-CD20 Fab′on Raji cell growth ;morphologic 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 f ragment Fab′,the apoptotic 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′.

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