Wang Jinghong, Boxer Linda M
Center for Molecular Biology in Medicine, Veterans Affairs Palo Alto Health Care System and the Department of Medicine, Stanford University School of Medicine, Stanford, California 94305, USA.
J Biol Chem. 2005 Apr 1;280(13):12766-73. doi: 10.1074/jbc.M412446200. Epub 2005 Feb 1.
Burkitt's lymphoma is invariably associated with chromosomal translocations that juxtapose the c-myc proto-oncogene with regulatory elements of the immunoglobulin heavy (IgH) or light chain loci resulting in the deregulation of c-myc expression. However, the enhancer elements mediating c-myc deregulation in vivo remain largely unidentified. To investigate the role of the IgH 3'-enhancers in c-myc deregulation, we used gene targeting to generate knock-in mice in which four DNase I hypersensitive regions from the murine IgH 3'-region were integrated into the 5'-region of the c-myc locus. The IgH 3'-enhancers induced the up-regulation of c-myc expression specifically in B cells of IgH-3'-E-myc mice. After approximately 10 months, the mice developed a Burkitt-like B cell lymphoma with the phenotype of B220+, IgM+, and IgD(low). Analysis of immunoglobulin gene rearrangements indicated that the lymphoma cells were of clonal origin. The presence of a rapidly expanding population of B cells in the spleen and bone marrow of young knock-in mice at 2-4 months of age was observed. Premalignant splenic B cells of knock-in mice showed higher spontaneous and induced apoptosis; however, malignant B cells were more resistant to apoptosis. The p53-ARF-Mdm2 pathway was disabled in half of the lymphomas examined, in most cases through Mdm2 overexpression. Although c-myc expression was increased in premalignant B cells, the promoter shift from P2 to P1 was observed only in malignant B cells. Our studies demonstrate that the IgH 3'-enhancers play an important role in c-myc deregulation and B cell lymphomagenesis in vivo.
伯基特淋巴瘤总是与染色体易位相关,这种易位使原癌基因c-myc与免疫球蛋白重链(IgH)或轻链基因座的调控元件并列,导致c-myc表达失调。然而,在体内介导c-myc失调的增强子元件在很大程度上仍未明确。为了研究IgH 3'-增强子在c-myc失调中的作用,我们利用基因靶向技术生成了基因敲入小鼠,其中来自小鼠IgH 3'-区域的四个DNase I高敏区域被整合到c-myc基因座的5'-区域。IgH 3'-增强子特异性地诱导了IgH-3'-E-myc小鼠B细胞中c-myc表达的上调。大约10个月后,这些小鼠发生了具有B220+、IgM+和IgD(低)表型的伯基特样B细胞淋巴瘤。免疫球蛋白基因重排分析表明淋巴瘤细胞是克隆起源的。在2-4月龄的年轻基因敲入小鼠的脾脏和骨髓中观察到存在快速扩增的B细胞群体。基因敲入小鼠的癌前脾脏B细胞显示出更高的自发和诱导凋亡;然而,恶性B细胞对凋亡更具抗性。在半数检测的淋巴瘤中,p53-ARF-Mdm2通路失活,在大多数情况下是通过Mdm2过表达。虽然在癌前B细胞中c-myc表达增加,但仅在恶性B细胞中观察到启动子从P2向P1的转变。我们的研究表明,IgH 3'-增强子在体内c-myc失调和B细胞淋巴瘤发生中起重要作用。