Belaud-Rotureau Marc-Antoine, Parrens Marie, Dubus Pierre, Garroste Jean-Christophe, de Mascarel Antoine, Merlio Jean-Philippe
Department of Histology and Molecular Pathology, Victor Segalen University, Bordeaux, France.
Mod Pathol. 2002 May;15(5):517-25. doi: 10.1038/modpathol.3880556.
Mantle cell lymphoma (MCL) diagnosis first relies on morphology and phenotype that may overlap with other B-cell lymphomas. Therefore, the demonstration of t(11;14)(q13;q32), the cytogenetic hallmark of MCL, is considered of diagnostic value. By studying a series of 35 MCL with characteristic morphology and phenotype (CD5+, CD10-, CD20+, CD23-), we have evaluated the applicability and the sensitivity of interphase fluorescence in situ hybridization (FISH) for t(11;14) detection and other techniques: (1) polymerase chain reaction (PCR) for amplification of t(11;14) genomic breakpoint, (2) competitive RT-PCR for the detection of cyclin D1 transcripts overexpression, and (3) immunohistochemistry (IHC) for cyclin D1 protein detection. Tissues from different origins were analyzed: lymph nodes (n = 24), spleen (n = 3), digestive biopsy (n = 3), tonsils (n = 3), and skin (n = 2). Interphase FISH was performed either on touch preparations (n = 11) and frozen (n = 9) or paraffin sections (n = 15). FISH analysis detected t(11;14) in 34/35 cases (97%) and demonstrated a recurrent CCND1 amplification in t(11;14)+ nuclei of the three blastoid MCL variants of our series. Genomic PCR analysis, hampered by the scattering of 11q13 breakpoints, was positive in only 13/35 cases (37%). RT-PCR analysis was applicable on nonepithelial tissues (27/35) and showed cyclin D1 transcript overexpression in all tested cases (27/35). IHC for cyclin D1 protein was performed either on frozen (n = 12) or on paraffin sections (n = 23), and its sensitivity was higher on paraffin sections (91%) than on frozen sections (25%). A cyclin D1 protein immunoreactivity was observed in 24/35 cases (69%). Our study emphasizes on the use of FISH analysis for the direct detection of t(11;14) because its applicability and sensitivity largely exceeded those of other techniques. It may also provide some informations on secondary cytogenetic changes of potential clinical relevance.
套细胞淋巴瘤（MCL）的诊断首先依赖于形态学和表型，而这些可能与其他B细胞淋巴瘤重叠。因此，t(11;14)(q13;q32)（MCL的细胞遗传学特征）的显示被认为具有诊断价值。通过研究一系列35例具有特征性形态学和表型（CD5+、CD10-、CD20+、CD23-）的MCL，我们评估了间期荧光原位杂交（FISH）检测t(11;14)以及其他技术的适用性和敏感性：（1）聚合酶链反应（PCR）扩增t(11;14)基因组断点，（2）竞争性逆转录PCR检测细胞周期蛋白D1转录本的过表达，以及（3）免疫组织化学（IHC）检测细胞周期蛋白D1蛋白。分析了来自不同来源的组织：淋巴结（n = 24）、脾脏（n = 3）、消化活检组织（n = 3）、扁桃体（n = 3）和皮肤（n = 2）。间期FISH在触片（n = 11）、冷冻切片（n = 9）或石蜡切片（n = 15）上进行。FISH分析在34/35例（97%）中检测到t(11;14)，并在我们系列中的三个母细胞样MCL变体的t(11;14)+细胞核中显示出复发性CCND1扩增。基因组PCR分析因11q13断点的分散而受阻，仅在13/35例（37%）中呈阳性。RT-PCR分析适用于非上皮组织（27/35），并在所有测试病例（27/35）中显示细胞周期蛋白D1转录本过表达。细胞周期蛋白D1蛋白的IHC在冷冻切片（n = 12）或石蜡切片（n = 23）上进行，其在石蜡切片上的敏感性（91%）高于冷冻切片（仅25%）。在24/35例（69%）中观察到细胞周期蛋白D1蛋白免疫反应性。我们的研究强调使用FISH分析直接检测t(11;14)，因为其适用性和敏感性大大超过了其他技术。它还可能提供一些关于潜在临床相关性的继发性细胞遗传学变化的信息。
