T细胞激活剂对Jurkat 细胞重组激活基因表达影响.docVIP

T细胞激活剂对Jurkat 细胞重组激活基因表达影响 　 作者：邹红云，马骊，姚新生，温茜，王小宁 【关键词】 T细胞激活剂,；Jurkat细胞；重组激活基因；逆转录聚合酶链反应 【Abstract】 AIM: To investigate the effects of T cell activators on the mRNA expression of recombinationactivating genes (RAGs) in Jurkat cells. METHODS: After Jurkat cells were treated with PHA (20 mg/L), antiCD3 mAb (1∶100), PMA(40 μg/L)+A23187 (0.5 μmol/L) for 6 and 12 h respectively, RAG1 and RAG2 mRNA were measured by RTPCR. Semiquantitative analysis was used to evaluate RAG1 and RAG2 mRNA levels in different groups. RESULTS: Compared with the control groups, the levels of RAG1 mRNA were significantly lower in different treated groups (Plt;0.05), and RAG1 mRNA levels were associated with treatment time. RAG2 mRNA levels in PHA treated groups were significantly lower than those in the control groups (Plt;0.05). No RAG2 mRNA was detected in PMA+A23187 treated groups. However, RAG2 mRNA level was significantly higher in antiCD3 mAb (12 h) treated group than that in control groups(Plt;0.05). CONCLUSION： RAG1 and RAG2 mRNA were coexpressed in Jurkat cells and could be regulated by T cell activators. The results may give a clue that Jurkat cells maybe provide an ideal cell line model for studying the regulation of RAGs in peripheral T cells. 【Keywords】 T cell activators; Jurkat cells; recombinationactivating genes; reverse transcriptase polymerase chain reaction 【摘要】 目的： 研究T细胞激活剂PHA，抗CD3 mAb, PMA+A23187对人T细胞白血病细胞株Jurkat重组激活基因（RAGs ）mRNA表达水平的影响. 方法： 采用RTPCR法检测不同T细胞激活剂作用不同时间后Jurkat 细胞中RAG1, RAG2 mRNA，以β肌动蛋白（βactin）的表达作为内参照进行灰度分析，比较不同组Jurkat细胞RAGs mRNA的表达水平 . 结果： 三种T细胞激活剂刺激诱导的Jurkat细胞RAG1 mRNA表达量明显低于对照组（Plt;0.05），且RAG1 mRNA表达量的下降与刺激诱导作用时间有关；PHA刺激诱导后RAG2表达量显著低于对照组（Plt;0.05）；PMA+A23187刺激诱导6, 12 h均未检测出RAG2 mRNA的表达； 而抗CD3 mAb作用12 h组RAG2 mRNA表达量显著高于对照组（Plt;0.05）. 结论： Jurkat细胞同时表达RAG1, RAG2 mRNA，并在一定诱导下RAGs表达量发生改变，因此有可能成为研究外周T细胞RAGs表达调节的一个潜在的细胞模型. 【关键词】 T细胞激活剂 ；Jurkat细胞；重组激活基因；逆转录聚合酶链反应 淋巴细胞特异性重组激活基因（recombination activating genes， RAG）1和RAG2是参与V(D)J重排和功能性抗原受体基因形成的一个首要条件［1］. 一般认为，RA