人T细胞白血病细胞株Jurkat的TCR基因重排  

T Cell Receptor Gene Recombination in Human T-cell Leukemia Cell Line Jurkat

作　　者：邹红云[1] 马骊[1] 罗微[1] 王小宁[2] 

机构地区：[1]南方医科大学生物技术学院分子免疫学研究所,广东广州510515 [2]华南理工大学生物科学与工程学院,广东广州510641

出　　处：《癌症》2006年第10期1198-1204,共7页Chinese Journal of Cancer

基　　金：国家重点基础研究发展规划"973"项目资助(No.2001CB510008)~~

摘　　要：背景与目的:近年的一些体外实验研究发现,重组激活基因(recombi-nationactivatinggene)编码的RAG1与RAG2(recombinationactivatinggenes,RAGs)蛋白能够介导DNA链的转位(transposition)作用,因而,可能与淋巴系统肿瘤性疾病的发生有关,但迄今尚未有明确定论。我们在实验中发现,代表T细胞发育成熟阶段的人白血病细胞株Jurkat同时表达重组激活基因RAG1和RAG2,并且经适当诱导后有RAGs表达的变化。本研究旨在确证Jurkat细胞是否发生RAGs介导的T细胞受体(T-cellreceptor,TCR)基因重排。方法:采用巢式和半巢式PCR检测T细胞受体D!-J!之间的信号结合T细胞受体删除DNA环(signaljointT-cellreceptorexcisionDNAcircles,sjTRECs);连接介导的PCR(LM-PCR)法检测TCRβ链位点重排中间物-重组信号序列(recombinationsignalsequence,RSS)断点;RT-PCR法检测V(D)J重排第二阶段非同源末端连接(non-homologousendjoining,NHEJ)途径中的核心蛋白Ku70/Ku80及末端脱氧核苷酸转移酶(terminaldeoxynucleotidyltransferase,TdT)。结果:在Jurkat细胞DNA中检测到结合区具有多样性特征的TCRDβ2-Jβ2sjTRECs和RSS5′端和3′端断裂点,并检测到TdT、Ku70/Ku80的表达。结论:Jurkat细胞有TCR基因重排的发生。Jurkat细胞可能成为研究RAGs和TCR基因重排与T细胞淋巴瘤的一个潜在的细胞模型。BACKGROUND ＆ OBJECTIVE： Recently, recombination activating gene （RAG）-mediated transposition has been found to contribute to chromosomal translocation and may be related to the occurrence of lymphoid malignancy; however, the underlying mechanism is unclear. Our previous study showed a co-expression of RAG1 and RAG2 in a human T-cell leukemia cell line Jurkat, which represents a mature stage of T cell development, and that the mRNA levels could be regulated by T cell activators. This study was to determine RAGs-mediated T cell receptor（TCR） gene recombination in Jurkat cells, and thus to provide a new thoughtway for studying the correlations of TCR gene recombination to lymphoid malignancy. METHODS-TCR DIS-Jβ signal joint T-cell receptor excision DNA circles （sjTRECs） were determined by nested and semi-nested polymerase chain reaction （PCR）. Double-strand DNA breaks at recombination signal sequences （RSSs） in the TCR β chain locus were detected by ligation-mediated polymerase chain reaction （LM-PCR）. TdT and Ku70/Ku80 were detected by reverse transcription-polymerase chain reaction （RT-PCR）. RESULTS： Characteristics of junctional diversity of Dβ2- Jβ2 sjTRECs and ds RSS breaks associated with Dβ2 5＇-site and Dβ2 3＇-site were detected in DNA of Jurkat cells. TdT and Ku70/KuSO were also detected. CONCLUSION： There is an ongoing TCR gene recombination in Jurkat cells. Jurkat may be used as an ideal cell line model for studying the correlation of TCR gene recombination to lymphoid malignancy.

关 键 词：白血病 JURKAT细胞 重组激活基因 TCR基因重排 

分 类 号：R346[医药卫生—基础医学]