Li J Y, Gaillard F, Moreau A, Harousseau J L, Laboisse C, Milpied N, Bataille R, Avet-Loiseau H
Laboratory of Hematology, Laboratory of Pathology, and Clinical Hematology Department, University Hospital, Nantes, France.
Am J Pathol. 1999 May;154(5):1449-52. doi: 10.1016/S0002-9440(10)65399-0.
To assess an unequivocal diagnosis of mantle cell lymphoma (MCL), we have developed a fluorescence in situ hybridization (FISH) assay, enabling the demonstration of t(11;14)(q13;q32) directly on pathological samples. We have first selected CCND1 and IGH probes encompassing the breakpoint regions on both chromosomes. Then, we have defined experimental conditions enabling us to obtain bright clear-cut signals in all of the samples, independently of the initial fixation conditions. We have analyzed single-cell suspensions from 26 formalin-fixed, paraffin-embedded MCL samples with this set of probes. In all cases, we have found a fusion signal (ie, a t(11;14)(q13;q32) translocation) in 14% to 99% of cells (median, 87%). So far, IGH-CCND1 fusions have been detected in all of the 51 MCL patients that we have analyzed by FISH (either on paraffin-embedded tumor samples or on peripheral blood samples). Regarding the low sensitivity of other techniques used to diagnose t(11;14)(q13;q32) (ie, 70% to 75% for cytogenetics and 50% to 60% for polymerase chain reaction), our FISH assay is by far the most sensitive technique. Moreover, because of the quality of the fluorescent signals and the rapidity of the experiment, this technique is widely applicable, even in routine cytogenetics or pathology laboratories. As MCL patients are usually refractory to standard therapy, an unambiguous diagnosis is needed to propose adapted therapeutic strategies, and this highly sensitive assay may be of great value for accurate diagnosis in difficult cases.
为评估套细胞淋巴瘤（MCL）的确切诊断，我们开发了一种荧光原位杂交（FISH）检测方法，能够直接在病理样本上显示t(11;14)(q13;q32)。我们首先选择了包含两条染色体断点区域的CCND1和IGH探针。然后，我们确定了实验条件，使我们能够在所有样本中获得明亮清晰的信号，而不受初始固定条件的影响。我们用这组探针分析了26个福尔马林固定、石蜡包埋的MCL样本的单细胞悬液。在所有病例中，我们在14%至99%的细胞中发现了融合信号（即t(11;14)(q13;q32)易位）（中位数为87%）。到目前为止，在我们通过FISH分析的所有51例MCL患者中（无论是石蜡包埋的肿瘤样本还是外周血样本）均检测到IGH-CCND1融合。鉴于用于诊断t(11;14)(q13;q32)的其他技术敏感性较低（即细胞遗传学为70%至75%，聚合酶链反应为50%至60%），我们的FISH检测方法是迄今为止最敏感的技术。此外，由于荧光信号质量高且实验快速，该技术广泛适用，即使在常规细胞遗传学或病理学实验室也是如此。由于MCL患者通常对标准治疗难治，因此需要明确的诊断以提出合适的治疗策略，而这种高灵敏度检测方法对于疑难病例的准确诊断可能具有重要价值。
