嵌合抗CD20抗体Fab′片段诱导Raji细胞凋亡过程中活性氧自由基的变化

目的　研究嵌合抗CD2 0抗体Fab′片段对B淋巴细胞株Raji细胞活性氧自由基 (ROS)的影响。 方法　利用MTT法测定Fab′片段抑制细胞生长 ,形态学方法和DNA琼脂糖凝胶电泳法观察Fab′片段对细胞凋亡的影响 ,用DCFH DA和双氢罗丹明 1 2 3(DHR )标记细胞内活性氧(ROS)变化 ,并用流式细胞仪检测。结果　MTT法测定抗CD2 0抗体Fab′片段对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 ,电镜观察 ,可见“凋亡小体”出现 ,DNA电泳可见典型的“梯形”条带 ,抗CD2 0抗体Fab′片段能增加Raji细胞内ROS水平 ,且随浓度增加而增加。 结论　ROS水平升高在抗CD2 0抗体Fab′片段诱导Raji细胞凋亡的过程中起到重要作用

ROS change in apoptosis of B lymphoma cells induced by chimeric anti-CD20 antibody fragment Fab′

AIM To study effect of chimeric anti CD20 antibody fragment Fab′ on ROS in B lymphoma Raji cells. METHODS MTT was used to observe the effect of anti CD20Fab on Raji cell growth; morphologic change through electron microscope and DNA ladder extracted were used to assay apoptosis induced by anti CD20Fab; ROS labeled with DCFH DA and 123 (DHR) in cells was processed on FACS. RESULTS The result of MTT indicated that the growth of Raji cells was inhibited by anti CD20Fab with dose dependently; the apoptotic bodies and DNA ladder were observed; ROS level in Raji cells was increased by anti CD20Fab. CONCLUSIONS The increase of ROS level is important in the apoptosis of Raji cells induced by anti CD20Fab.