MLL Munich Leukemia Laboratory, Munich, Germany.
Exp Hematol. 2013 Dec;41(12):1028-37. doi: 10.1016/j.exphem.2013.09.004. Epub 2013 Sep 20.
Molecular diagnosis of mantle cell lymphoma (MCL) can be difficult because the t(11;14)/IGH@-CCND1 is extremely heterogeneous at the DNA level. Aiming to establish a reliable molecular tool that could be easily implemented in routine diagnostics, we developed a new real-time polymerase chain reaction (PCR) assay for CCND1 expression measurement and evaluated 451 cases: 142 MCL, 76 chronic lymphocytic leukemia, 20 hairy cell leukemia, 13 hairy cell leukemia-variant, 20 splenic marginal zone lymphoma, 91 other mature B-cell neoplasms, 29 other hematologic neoplasms, and 60 healthy individuals. Sensitivity of the real-time PCR assay was up to 10(-4). In t(11;14)/IGH@-CCND1 positive lymphoma samples (n = 150), median %CCND1/ABL1 expression level was 178.2 (range: 1.5-4, 152.0). Normalized by t(11;14)/IGH@-CCND1 positive cells as determined by fluorescence in situ hybridization IGH@-CCND1 positive samples showed a median %CCND1/ABL1 of 445.8 (range: 17.9-4,848.5). A normalized %CCND1/ABL1 expression of at least 17.0 was chosen as threshold for CCND1 positivity. For unnormalized samples, the positive detection rate of t(11;14)/IGH@-CCND1 by CCND1 expression was 87.3%. Healthy individuals had low %CCND1/ABL1 (median, 1.1; range, 0.0-7.8). The negative predictive value for exclusion of a t(11;14)/IGH@-CCND1 by CCND1 expression was 95.3% by the above threshold. %CCND1/ABL1 was higher in MCL than in the remaining B-cell lymphomas (mean ± SD, 392.9 ± 685.3 vs. 46.0 ± 305.0; p < 0.001). In 66 follow-up samples, CCND1 showed 2.5-3.5 log reduction after chemotherapy and increase at relapse. CCND1 expression could serve as adjunct to other techniques in diagnosis and follow-up of B-cell lymphomas.
套细胞淋巴瘤（MCL）的分子诊断可能具有挑战性，因为 t（11；14）/IGH@-CCND1 在 DNA 水平上存在极大的异质性。为了建立一种可靠的分子工具，使其易于在常规诊断中实施，我们开发了一种新的实时聚合酶链反应（PCR）检测 CCND1 表达的方法，并评估了 451 例病例：142 例 MCL、76 例慢性淋巴细胞白血病、20 例毛细胞白血病、13 例毛细胞白血病变异型、20 例脾边缘区淋巴瘤、91 例其他成熟 B 细胞肿瘤、29 例其他血液系统肿瘤和 60 例健康个体。实时 PCR 检测的灵敏度高达 10(-4)。在 t（11；14）/IGH@-CCND1 阳性淋巴瘤样本（n=150）中，CCND1/ABL1 表达水平中位数为 178.2（范围：1.5-4，152.0）。通过荧光原位杂交 IGH@-CCND1 阳性样本确定的 t（11；14）/IGH@-CCND1 阳性细胞进行归一化后，CCND1/ABL1 的中位数为 445.8（范围：17.9-4，848.5）。选择至少 17.0 的归一化 CCND1/ABL1 表达作为 CCND1 阳性的阈值。对于未归一化的样本，通过 CCND1 表达检测 t（11；14）/IGH@-CCND1 的阳性检出率为 87.3%。健康个体的 CCND1/ABL1 水平较低（中位数，1.1；范围，0.0-7.8）。通过上述阈值，排除 t（11；14）/IGH@-CCND1 的阴性预测值为 95.3%。CCND1 在 MCL 中的表达高于其他 B 细胞淋巴瘤（均值±标准差，392.9±685.3 与 46.0±305.0；p<0.001）。在 66 例随访样本中，化疗后 CCND1 表达降低 2.5-3.5 个对数级，复发时升高。CCND1 表达可作为其他技术在 B 细胞淋巴瘤诊断和随访中的辅助手段。
