【摘要】 目的研究抗CD20嵌合抗体片段Fab′突变体诱导B淋巴细胞凋亡的发生、凋亡相关基因表达以及细胞内钙离子的浓度变化。方法以人B淋巴细胞(Raji细胞)为凋亡细胞模型,通过MTT法、Western blot和流式细胞仪等方法观察bcl-2/bax基因表达、细胞色素c及胞内Ca2+的变化。结果抗CD20嵌合抗体片段Fab′突变体对Raji细胞生长增殖具有明显的抑制作用,并呈剂量依赖性。细胞中bcl-2表达有微弱的下降,而bax表达明显升高,并出现细胞色素c的释放和胞内Ca2+浓度的升高。结论抗CD20嵌合抗体片段Fab′突变体诱导Raji细胞凋亡,与bcl-2/bax表达、细胞色素c的释放和胞内Ca2+浓度的升高有关。更多还原

【Abstract】 Objective:To investigate the effect of chimeric anti-CD20 antibody Fab′fragment on apoptosis, apoptosis related genes expression, and intracellular Ca 2+ concentration of B lymphoma cells. Methods: The proliferation of human B lymphoma cell line Raji was measured by MTT assay. The release of cytochrome c and expression of bcl-2/bax was detected by Western blotting. Apoptosis of Raji cells was determined by FACS analysis. Intracellular calcium concentration was measured by fluorescence method. Results:Chimeric anti-CD20 antibody Fab′fragment inhibited proliferation of Raji cells in a dose-dependent!manner, slightly down-regulated bcl-2 protein expression, but significantly up-regulated bax expression. Cytochrome c was released and intracellular Ca 2+ concentration was increased. Conclusion:Chimeric anti-CD20 antibody Fab′ fragment induced apoptosis of Raji cells which was related with changes of bcl-2/bax, release of cytochrome c, and increase in intracellular Ca 2+ concentration.更多还原