Department of Clinical Cytology and Cytogenetics, Nîmes University Hospital, Nîmes, France.
Mol Cancer Ther. 2010 Nov;9(11):3083-9. doi: 10.1158/1535-7163.MCT-10-0595. Epub 2010 Nov 2.
Chronic myeloid leukemia (CML) patients treated with imatinib develop frequent resistance generally due to a point mutation. Recently, large rearrangements of abl sequence have also been described. In this study, we focused on the complete deletion of exon 7. We screened for bcr-abl(delexon7) in 63 resistant patients by high-resolution melting (HRM) analysis and direct sequencing. Moreover, we analyzed expression of abl(delexon7) and bcr-abl(delexon7) in 17 CML patients at diagnosis, 32 patients at resistance, and 20 negative controls by quantitative PCR or fragment length analysis. bcr-abl(delexon7) was detected on 34 (54%) among 63 resistant patients by HRM, showing an increase in the sensitivity of screening, because only 3.2% could be detected by direct sequencing. This deletion was not associated with a point mutation (P = 0.3362). In addition, abl(delexon7) was found in all tested samples with the same pattern of expression, suggesting an alternative splicing mechanism. In the bcr-abl component, there was no statistical difference between CML patients at diagnosis and resistant patients (P = 0.2815) as regarding bcr-abl(delexon7) proportion, thus arguing against involvement of deletion in resistance. Moreover, among two patients harboring bcr-abl(delexon7) at diagnosis, one experienced a complete disappearance of this transcript, and the other decreased >75% at resistance. In conclusion, bcr-abl(delexon7) is frequently observed in CML patients when using sensitive techniques. It seems to be the result of an alternative splicing mechanism and to be independent from the occurrence of resistance.
慢性髓性白血病 (CML) 患者在接受伊马替尼治疗后会因点突变而经常产生耐药性。最近,也描述了 abl 序列的大型重排。在这项研究中,我们重点研究了外显子 7 的完全缺失。我们通过高分辨率熔解 (HRM) 分析和直接测序筛选了 63 名耐药患者中的 bcr-abl(delexon7)。此外,我们通过定量 PCR 或片段长度分析分析了 17 名 CML 患者在诊断时、32 名在耐药时和 20 名阴性对照者中的 abl(delexon7)和 bcr-abl(delexon7)的表达。通过 HRM 在 63 名耐药患者中的 34 名 (54%) 检测到 bcr-abl(delexon7),这提高了筛查的灵敏度,因为直接测序只能检测到 3.2%。这种缺失与点突变无关 (P = 0.3362)。此外,在所有检测的样本中都发现了 abl(delexon7),表达模式相同,提示存在替代剪接机制。在 bcr-abl 成分中,在诊断时和耐药患者之间, bcr-abl(delexon7)的比例没有统计学差异 (P = 0.2815),因此不支持缺失与耐药有关。此外,在两名诊断时携带 bcr-abl(delexon7)的患者中,一名患者这种转录本完全消失,另一名患者在耐药时减少了 >75%。总之,在使用敏感技术时, CML 患者中经常观察到 bcr-abl(delexon7)。它似乎是一种替代剪接机制的结果,与耐药的发生无关。