CD3ε和5-氟尿嘧啶在T淋巴细胞凋亡中的协同作用及P53的表达  

Synergetic role of CD3ε and 5-fluorouracil in T Lymphocyte apoptosis and P53 expression

作　　者：程宏[1] 刘彦信[1] 张锦春[1] 刘士廉[1] 郑德先[1] 

机构地区：[1]中国协和医科大学基础医学院中国医学科学院基础医学研究所医学分子生物学国家重点实验室,北京100005

出　　处：《中华医学杂志》2003年第22期1962-1967,共6页National Medical Journal of China

基　　金：国家重大基础性研究发展规划资助项目(G19990 5 3 90 5 );国家自然科学基金资助项目 ( 3 0 170 3 62 )

摘　　要：目的　研究CD3ε分子与化疗药物 5 氟尿嘧啶 (5 FU)在T淋巴细胞凋亡中的协同作用及P5 3的表达 ,为探索T淋巴细胞凋亡的分子机制打下基础 ,为相关肿瘤的治疗提供新的实验依据。方法　培养人T淋巴白血病细胞株JK ,转染了突变的CD3εITAM的TJK及T3JK细胞株。用CD3ε特异的单克隆抗体 (2 0 0 μg/ml)和 /或 5 FU(2 5 μg/ml)刺激不同的T淋巴细胞诱导细胞凋亡 ,用噻唑蓝比色法检测细胞存活率 ,并计算细胞死亡率 ;用瞬时转染野生型或突变型P5 3表达质粒的方法和Western印迹检测P5 3蛋白质的表达 ,分析其在T淋巴细胞凋亡中的作用。结果　在CD3ε和 5 FU分别诱导T淋巴细胞凋亡过程中 ,均伴随着P5 3蛋白质表达增加 ;两者联合使用使T淋巴细胞凋亡和P5 3蛋白质表达显著增加 ;P5 3过表达可明显增加T淋巴细胞对 5 FU的敏感性。结论　抗CD3ε抗体和 5 FU联合作用可显著提高P5 3蛋白质表达 ,两者在T淋巴细胞凋亡中具有协同效应。Objective To investigate the relationship between apoptosis induced by CD3ε and 5-fluorouracil (5-FU), and study the P53 expression in the apoptosis process provide a novel insight and useful information of the apoptosis signaling pathway induced by CD3ε and/or 5-FU, and an important implication for the treatment of T-lymphocyte leukemia. Methods The viabilities of Jurkat T lymphocytes (JK), TJK [JK with over-expression of the CD8ε chimeria molecule] and T3JK [JK with over-expression of the CD8ε(Y170F/Y181F) mutation molecule] cells were cultmed and treated with pre-coated anti-CD8 mAb (200 μg/ml) and/or 5-FU (2.5 μg/ml) were detected with MTS assay and the apoptosis percentages were calculated. Western blot was used to detect P53 expression. To confirm the role of P53 in 5-FU-treated T lymphocytes, pCMV-p53 plasmid with wild type or mutant p53 were co-transfected transiently with pEGFP-c1 into TJK and T3JK cells, respectively. Results CD3ε or 5-FU induced apoptosis of TJK with increase of P53 expression. Co-treatment with CD3ε specific antibody and 5-FU elevated the apoptotic rates and P53 expression in TJK cells remarkably. The cells transfected with wild-type p53 exhibited more sensitivity to 5-FU than that transfected with mutant p53. Conclusion Co-treatment of CD3ε and 5-FU increases the apoptosis and p53 expression, suggesting that there is a synergetic role of CD3ε and 5-FU on T lymphocytes.

关 键 词：CD3ε 5-氟尿嘧啶 T淋巴细胞凋亡 协同作用 P53 表达 单克隆抗体 

分 类 号：R392.1[医药卫生—免疫学]