Rossi Michael R, Laduca Jeff, Cowell John K, Srivastava Bejai I S, Matsui Seiichi
SKY Core Facility, Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, NY 14263, United States.
Leuk Res. 2008 Mar;32(3):455-63. doi: 10.1016/j.leukres.2007.04.019. Epub 2007 Jul 20.
We performed aCGH, SKY/FISH, molecular mapping and expression analyses on a permanent CD8+ NK/T cell line, 'SRIK-NKL' established from a lymphoma (ALL) patient, in attempt to define the fundamental genetic profile of its unique NK phenotypes. aCGH revealed hemizygous deletion of 6p containing genes responsible for hematopoietic functions. The SKY demonstrated that a constitutive reciprocal translocation, rcpt(5;14)(p13.2;q11) is a stable marker. Using somatic hybrids containing der(5) derived from SRIK-NKL, we found that the breakpoint in one homologue of no. 5 is located upstream of IL7R and also that the breakpoint in no. 14 is located within TRA@. The FISH analysis using a BAC which contains TRA@ and its flanking region further revealed a approximately 231kb deletion within 14q11 in the der(5) but not in the normal homologue of no. 14. The RT-PCR analysis detected mRNA for TRA@ transcripts which were extending across, but not including, the deleted region. IL7R was detected at least at mRNA levels. These findings were consistent with the immunological findings that TRA@ and IL7R are both expressed at mRNA levels and TRA@ at cytoplasmic protein levels in SRIK-NKL cells. In addition to rcpt(5;14), aCGH identified novel copy number abnormalities suggesting that the unique phenotype of the SRIK-NKL cell line is not solely due to the TRA@ rearrangement. These findings provide supportive evidence for the notion that SRIK-NKL cells may be useful for studying not only the function of NK cells but also genetic deregulations associated with leukemiogenesis.
我们对从一名淋巴瘤(ALL)患者建立的永久性CD8 + NK/T细胞系“SRIK-NKL”进行了比较基因组杂交(aCGH)、光谱核型分析(SKY)/荧光原位杂交(FISH)、分子定位和表达分析,试图确定其独特NK表型的基本遗传特征。aCGH显示6p上负责造血功能的基因半合子缺失。SKY表明,一种组成性相互易位rcpt(5;14)(p13.2;q11)是一个稳定标记。利用含有源自SRIK-NKL的der(5)的体细胞杂种,我们发现5号染色体一个同源物中的断点位于IL7R上游,14号染色体中的断点位于TRA@内。使用包含TRA@及其侧翼区域的细菌人工染色体(BAC)进行的FISH分析进一步显示,der(5)中14q11内有大约231kb的缺失,而14号染色体的正常同源物中没有。逆转录聚合酶链反应(RT-PCR)分析检测到TRA@转录本的mRNA,其延伸穿过但不包括缺失区域。至少在mRNA水平检测到了IL7R。这些发现与免疫结果一致,即SRIK-NKL细胞中TRA@和IL7R在mRNA水平均有表达,TRA@在细胞质蛋白水平有表达。除了rcpt(5;14),aCGH还发现了新的拷贝数异常,表明SRIK-NKL细胞系的独特表型不仅仅是由于TRA@重排。这些发现为SRIK-NKL细胞不仅可用于研究NK细胞功能,还可用于研究与白血病发生相关的基因失调这一观点提供了支持性证据。