Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas and Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid, Spain.
PLoS One. 2010 Dec 28;5(12):e15525. doi: 10.1371/journal.pone.0015525.
Chronic myeloid leukemia (CML) is a malignant clonal disorder of the hematopoietic system caused by the expression of the BCR/ABL fusion oncogene. Although it is well known that CML cells are genetically unstable, the mechanisms accounting for this genomic instability are still poorly understood. Because the Fanconi anemia (FA) pathway is believed to control several mechanisms of DNA repair, we investigated whether this pathway was disrupted in CML cells. Our data show that CML cells have a defective capacity to generate FANCD2 nuclear foci, either in dividing cells or after DNA damage. Similarly, human cord blood CD34(+) cells transduced with BCR/ABL retroviral vectors showed impaired FANCD2 foci formation, whereas FANCD2 monoubiquitination in these cells was unaffected. Soon after the transduction of CD34(+) cells with BCR/ABL retroviral vectors a high proportion of cells with supernumerary centrosomes was observed. Similarly, BCR/ABL induced a high proportion of chromosomal abnormalities, while mediated a cell survival advantage after exposure to DNA cross-linking agents. Significantly, both the impaired formation of FANCD2 nuclear foci, and also the predisposition of BCR/ABL cells to develop centrosomal and chromosomal aberrations were reverted by the ectopic expression of BRCA1. Taken together, our data show for the first time a disruption of the FA/BRCA pathway in BCR/ABL cells, suggesting that this defective pathway should play an important role in the genomic instability of CML by the co-occurrence of centrosomal amplification and DNA repair deficiencies.
慢性髓性白血病(CML)是一种造血系统的恶性克隆性疾病,由 BCR/ABL 融合癌基因的表达引起。虽然众所周知 CML 细胞的遗传不稳定,但导致这种基因组不稳定的机制仍知之甚少。由于范可尼贫血(FA)途径被认为可以控制几种 DNA 修复机制,因此我们研究了该途径是否在 CML 细胞中受到破坏。我们的数据表明,CML 细胞在有丝分裂细胞或 DNA 损伤后,生成 FANCD2 核斑的能力有缺陷。同样,转导了 BCR/ABL 逆转录病毒载体的人脐带血 CD34+细胞形成 FANCD2 焦点的能力受损,而这些细胞中的 FANCD2 单泛素化不受影响。在将 CD34+细胞转导 BCR/ABL 逆转录病毒载体后不久,就观察到大量具有多余中心体的细胞。同样,BCR/ABL 诱导了大量的染色体异常,同时在暴露于 DNA 交联剂后介导了细胞存活优势。重要的是,FANCD2 核斑形成受损,以及 BCR/ABL 细胞易发生中心体和染色体异常的倾向,都可以通过 BRCA1 的异位表达得到逆转。总之,我们的数据首次显示 FA/BRCA 途径在 BCR/ABL 细胞中受到破坏,表明该缺陷途径通过中心体扩增和 DNA 修复缺陷的共同发生,在 CML 的基因组不稳定性中应发挥重要作用。