An interphase fluorescence in situ hybridisation assay for the detection of 3q26.2/EVI1 rearrangements in myeloid malignancies
An interphase fluorescence in situ hybridisation assay for the detection of 3q26.2/EVI1 rearrangements in myeloid malignancies
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DOI：
10.1111/j.1365-2141.2007.06505.x
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发表时间：
2007-03-01
影响因子：
6.5
通讯作者：
Slovak, Marilyn L.
中科院分区：
文献类型：
--
作者：
Bobadilla, Dolores;Enriquez, Elmon L.;Slovak, Marilyn L.
涉及带3q26.2的染色体重排与髓系恶性肿瘤、人类嗜亲病毒整合位点-1（EVI 1）基因的异常表达、不良预后和侵袭性临床病程相关。3q26.2重排的特征是异质性，通常难以在质量差的中期细胞中检测到。为了开发用于检测3q26.2/EVI 1畸变的双色荧光原位杂交（FISH）测定，系统地评价了对应于EVI 1基因区域的一系列10个BAC克隆，并将其缩小到两个探针组;一个探针组包括延伸着丝粒的EVI 1基因，而第二个探针组覆盖EVI 1基因并延伸端粒。在不同处理时间点收集的35份细胞遗传学定义的3q26.2重排患者样本、inv（3）（q21q26.2）Kasumi-4细胞系和10份已知阴性样本上评价了两种探针集。双探针组策略鉴别了所有样品，尽管观察到了巨大的断点异质性。在急性髓性白血病和骨髓增生异常综合征病例的样本中，大多数倒位断裂点位于EVI 1的3'端，而3q26.2易位断裂点经常定位于EVI 1的5'端。然而，两个3q26.2易位样品在EVI 1的3'端具有断裂点。大多数inv（3q）慢性粒细胞白血病样本显示EVI 1基因内的断点。这项研究表明，尽管观察到3q26.2畸变存在广泛的断点异质性，但该FISH策略可有效检测骨髓恶性肿瘤中的3q26.2异常。
Chromosome rearrangements involving band 3q26.2 are associated with myeloid malignancies, aberrant expression of the human ecotropic virus integration site-1 (EVI1) gene, an unfavourable prognosis and an aggressive clinical course. The 3q26.2 rearrangements are characteristically heterogeneous and typically difficult to detect in poor quality metaphases. To develop a dual-colour fluorescence in situ hybridisation (FISH) assay for the detection of 3q26.2/EVI1 aberrations, a series of 10 BAC clones corresponding to the EVI1 gene region were systematically evaluated and narrowed down to two probe sets; one probe set encompassed the EVI1 gene extending centromeric, while the second probe set covered the EVI1 gene and extends telomeric. Both probe sets were evaluated on 35 patient samples with cytogenetically defined 3q26.2 rearrangements collected at various treatment time points, the inv(3)(q21q26.2) Kasumi-4 cell line, and 10 known negative samples. The two-probe set strategy identified all samples, despite the vast breakpoint heterogeneity observed. In samples from acute myeloid leukaemia and myelodysplastic syndrome cases, the majority of inversion breakpoints were 3' to EVI1 whereas 3q26.2 translocation breakpoints frequently mapped 5' to EVI1. However, two 3q26.2 translocation samples had breakpoints 3' to EVI1. Most inv(3q) chronic myeloid leukaemia samples showed breakpoints within the EVI1 gene. This study demonstrated that, despite the extensive breakpoint heterogeneity observed with 3q26.2 aberrations, this FISH strategy is effective for the detection of 3q26.2 abnormalities in myeloid malignancies.