Müller J R, Potter M, Janz S
Laboratory of Genetics, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12066-70. doi: 10.1073/pnas.91.25.12066.
The translocation of c-myc on chromosome (chr.) 15 to an immunoglobulin heavy-chain switch region on chr. 12 is the critical oncogenic step in pristane-induced plasmacytoma (PCT) development in BALB/cAnPt mice. Applying a recently developed PCR method, we have been able to detect the most commonly occurring illegitimate recombinations between alpha-chain switch region (S alpha) and c-myc in preneoplastic B cells residing in mesenteric oil granuloma (OG) tissues 7-30 days postpristane. In this study, we compare the nucleotide sequences at the S alpha/c-myc breaksites on both the c-myc-activating chr. 12+ and the reciprocal chr. 15- from eight transplanted PCTs, seven primary PCTs, and five OGs that contained six B-cell clones. These junction sequences revealed a remarkable diversity of S alpha/c-myc recombinations. In nine cases--four PCTs and five B-cell clones--nearly precise reciprocal exchanges with a loss of only 3-35 bp in c-myc were found. Large deletions in c-myc that removed 369-878 bp were observed in seven PCTs but not in early B cells. Duplications of c-myc ranging from 103 to 229 bp were also restricted to PCTs and noticed in four cases. Clonally related but different reciprocal recombinations, 38 bp apart on chr. 12+ and 15 bp apart on chr. 15-, were isolated from two different specimens of the same OG tissue from a BALB/c mouse 30 days postpristane. A second OG from another 30-day mouse yielded four recombinational fragments--two clonally related chr. 12(+)-specific fragments and two chr. 15(-)-specific fragments--one of which carried a 143-bp insertion of a microsatellite at the breaksite. We suggest that the initial recombinational break-point regions between S alpha and c-myc in plasmacytoma precursor cells at the time of immunoglobulin heavy-chain switching are intrinsically labile and characterized by a persisting instability of c-myc, which can result in large secondary deletions of c-myc.
在BALB/cAnPt小鼠中,15号染色体上的c-myc易位至12号染色体上的免疫球蛋白重链转换区是 pristane诱导的浆细胞瘤(PCT)发生过程中的关键致癌步骤。应用最近开发的PCR方法,我们能够检测到在pristane注射后7 - 30天,肠系膜油肉芽肿(OG)组织中癌前B细胞内α链转换区(Sα)与c-myc之间最常见的异常重组。在本研究中,我们比较了来自8个移植性PCT、7个原发性PCT以及5个含有6个B细胞克隆的OG的c-myc激活的12号染色体(chr. 12+)和对应的15号染色体(chr. 15-)上Sα/c-myc断裂位点的核苷酸序列。这些连接序列显示出Sα/c-myc重组具有显著的多样性。在9个病例中——4个PCT和5个B细胞克隆——发现了几乎精确的相互交换,c-myc中仅缺失3 - 35 bp。在7个PCT中观察到c-myc中有369 - 878 bp的大片段缺失,但在早期B细胞中未观察到。c-myc的103至229 bp的重复也仅限于PCT,在4个病例中被发现。从一只BALB/c小鼠pristane注射后30天的同一OG组织的两个不同标本中分离出克隆相关但不同的相互重组,在chr. 12+上相距38 bp,在chr. 15-上相距15 bp。另一只30天龄小鼠的第二个OG产生了四个重组片段——两个克隆相关的chr. 12(+)-特异性片段和两个chr. 15(-)-特异性片段——其中一个在断裂位点携带了一个143 bp的微卫星插入。我们认为,在免疫球蛋白重链转换时,浆细胞瘤前体细胞中Sα与c-myc之间的初始重组断点区域本质上是不稳定的,其特征是c-myc持续不稳定,这可能导致c-myc的大量继发性缺失。