T cell receptor gene recombination in human T-cell leukemia cell line jurkat

• Published in: Ai zheng = Aizheng = Chinese journal of cancer Vol. 25; no. 10; p. 1198
• Main Authors: Zou, Hong-Yun, Ma, Li, Luo, Wei, Wang, Xiao-Ning
• Format: Journal Article
• Language: Chinese
• Published: China 01.10.2006
• Subjects: Antigens, Nuclear - metabolism; Base Sequence; DNA - genetics; DNA Breaks, Double-Stranded; DNA Nucleotidylexotransferase - metabolism; DNA-Binding Proteins - metabolism; Genes, T-Cell Receptor; Humans; Jurkat Cells; Ku Autoantigen; Leukemia, T-Cell - genetics; Molecular Sequence Data; Receptors, Antigen, T-Cell, alpha-beta - genetics; Recombination, Genetic

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. TCR Dbeta-Jbeta 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 beta chain locus were detected by ligation-mediated polymerase chain reaction (LM-PCR). TdT and Ku70/Ku80 were detected b