Molecular analysis of TCRB and ABL in a t(7;9)-containing cell line (SUP-T3) from a human T-cell leukemia.
Molecular analysis of TCRB and ABL in a t(7;9)-containing cell line (SUP-T3) from a human T-cell leukemia.
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对来自人 T 细胞白血病的含 t(7;9) 细胞系 (SUP-T3) 中的 TCRB 和 ABL 进行分子分析。
DOI：
10.1073/pnas.84.1.251
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发表时间：
1987
影响因子：
11.1
通讯作者：
Diaz,MO
中科院分区：
文献类型：
--
作者：
Westbrook,CA;Rubin,CM;LeBeau,MM;Kaminer,LS;Smith,SD;Rowley,JD;Diaz,MO
7 号和 9 号染色体之间的易位，即 t(7;9)，已在源自患有急性 T 细胞淋巴细胞白血病或淋巴瘤的儿童恶性细胞的细胞系中得到描述。我们对这样一种细胞系 SUP-T3 的细胞遗传学分析表明，7 号和 9 号染色体上的断点分别位于 q36 和 q34 条带内，对应于编码 T 细胞受体 TCRB β 链的基因以及与 Abelson 鼠白血病病毒转化基因 ABL 同源的基因的位置。我们研究了这些基因在 t(7;9) 中的作用。 TCRB 和 ABL 探针与 SUP-T3 中期细胞的原位染色体杂交表明，ABL 从 9 号染色体易位到 7 号染色体，并且全部或部分 TCRB 从 7 号染色体易位到 9 号染色体。Southern 印迹分析显示，两个 TCRB 等位基因都发生了重排；然而，无法确定易位断点是否位于该基因内。使用脉冲场凝胶电泳和 Southern blot 分析来检查 ABL 基因座的 500 多个碱基；我们得出的结论是，与 v-abl 同源的序列在 250 kb 范围内没有任何方向的重排。此外，体外磷酸化测定中未检测到异常 ABL 蛋白。这些结果表明，在 SUP-T3 中，9 号染色体上的断点位于 ABL 附近，并且该断点不会导致 ABL 蛋白发生明显改变。因此，我们假设 9 号染色体 q34 带上的另一个基因在这种易位中发挥了作用。这项研究还表明，脉冲场凝胶电泳是分析人类染色体易位的强大新工具。
A translocation between chromosomes 7 and 9, t(7;9), has been described in cell lines derived from the malignant cells of children with acute T-cell lymphoblastic leukemia or lymphoma. Our cytogenetic analysis of one such cell line, SUP-T3, demonstrates that the breakpoints on chromosomes 7 and 9 lie within bands q36 and q34, respectively, corresponding to the location of the gene encoding the beta chain of the T-cell receptor, TCRB, and the gene homologous to the transforming gene of the Abelson murine leukemia virus, ABL. We investigated the role of these genes in the t(7;9). In situ chromosomal hybridization of TCRB and ABL probes to metaphase cells from SUP-T3 demonstrated that ABL is translocated from chromosome 9 to 7 and that all or part of TCRB is translocated from chromosome 7 to 9. Southern blot analysis revealed that both TCRB alleles were rearranged; however, it could not be determined whether the translocation breakpoint lies within this gene. Pulsed-field gel electrophoresis and Southern blot analysis were used to examine more than 500 kilobases of the ABL locus; we concluded that there are no rearrangements within 250 kb in either direction of the sequences homologous to v-abl. Additionally, no abnormal ABL protein was detected in an in vitro phosphorylation assay. These results indicate that, in SUP-T3, the breakpoint on chromosome 9 lies proximal to ABL and that the break results in no apparent alteration of the ABL protein. We therefore hypothesize that another gene on chromosome 9, at band q34, plays a role in this translocation. This study also demonstrates that pulsed-field gel electrophoresis is a powerful new tool for the analysis of human chromosomal translocations.