酪氨酸磷酸化抑制剂AG490抑制Jurkat T细胞增殖  

Inhibitor of tyrosine phosphorylation AG490 inhibits the proliferation of Jurkat T cells

作　　者：毛成全[1] 邢飞跃[1] 郭中峰[1] 方志远[1] 

机构地区：[1]暨南大学生命科学技术学院组织移植与免疫中心,广东广州510632

出　　处：《细胞与分子免疫学杂志》2010年第3期214-217,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30471635);生物化学与分子生物学广东省重点学科资助(2009)

摘　　要：目的:通过AG490对Jurkat T细胞活化、增殖、周期、凋亡及ICBP90蛋白表达的影响,探讨阻断JAK/STAT信号通路以抑制Jurkat T细胞生长的可能性及其初步机制。方法:以Jurkat T细胞为模型,应用双荧光抗体标记结合流式细胞仪检测AG490对Jurkat T细胞表面分子CD69和CD25表达的影响;利用噻唑蓝(MTT)比色法观察AG490对Jurkat T细胞增殖的影响;采用碘化丙锭(PI)染色检测AG490对Jur-kat T细胞周期的影响;应用Annexin V-FITC和PI双染色检测AG490对Jurkat T细胞凋亡的影响;Western blot检测AG490对Jurkat T细胞中ICBP90蛋白表达的影响以确定其与AG490抑制Jurkat T细胞增殖的关系。结果:随着AG490浓度从1 mmol/L增至30 mmol/L,细胞停滞于G0/G1期,阻止其进入S期和G2/M期,导致Jurkat T细胞ICBP90蛋白的表达显著降低;AG490对细胞的抑制作用于24 h最为明显,抑制率可达27.37%,呈剂量依赖关系;AG490不能明显抑制Jurkat T细胞的活化或促进其凋亡。结论:AG490能明显抑制Jurkat T细胞的生长,其抑制作用可能通过下调Jurkat T细胞ICBP90蛋白的表达与细胞周期阻滞有关,而不是通过促进细胞凋亡而实现。AIM：To study effect of AG490 on the activation,proliferation,cell cycle,apoptosis and the expression of protein ICBP90 of Jurkat T cells.To explore the possibility of inhibiting and the initial growth mechanism of Jurkat T cells by blocking JAK signaling pathway.METHODS：The expression level of CD69 and CD25 on the surface of Jurkat T cells was evaluated by fluorescin-conjugated monoclonal antibody double labeling technique.The proliferation of Jurkat T cells was analyzed by MTT method.The cycle of Jurkat T cell was analyzed by PI staining and Flow Cytometry.The apoptosis of Jurkat T cells was analyzed by Annexin V-FITC and PI staining.The expression of ICBP90 that correlated with the cell proliferation was checked by Western blot.RESULTS：The results came out that the proliferation capacity of Jurkat T cells was decreased with the increasing of AG490 ranging from 1 mmol/L to 30 mmol/L in a dose-dependent manner with the highest inhibitory rate of 27.37%.Cell-cycle analysis revealed that AG490 arrest it in G0/G1 phase and inhibited its entrance into S and G2/M phase,and reduced the expression of protein ICBP90,especially in the 24 h treatment.But the apoptosis of Jurkat T cells was not promoted by AG490.CONCLUSION：AG490 significantly inhibited the growth of Jurkat T cells,and its inhibition may reduce the expression of protein ICBP90,which was related to the cell cycle arrest rather than apoptosis.

关 键 词：AG490 JURKAT T细胞 增殖 周期 ICBP90 

分 类 号：R392.12[医药卫生—免疫学]