Thomázy Vilmos A, Luthra Rajyalakshmi, Uthman Margaret O, Davies Peter J A, Medeiros L Jeffrey
Department of Pathology and Laboratory Medicine, The University of Texas-Houston Medical School, Houston, USA.
J Mol Diagn. 2002 Nov;4(4):201-8. doi: 10.1016/S1525-1578(10)60704-0.
Cyclin D1 overexpression is a valuable marker for the diagnosis of mantle cell lymphoma (MCL). We used a real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) method to quantify levels of cyclin D1, CD20, and cyclophilin A mRNA in manually microdissected, paraffin-embedded tissue sections using an ABI 7700 qRT-PCR system. The study group included 21 cases of MCL and 37 cases of other types of B-cell non-Hodgkin's lymphoma. Cyclin D1 mRNA copy number was normalized to CD20 and cyclophilin A mRNA and evaluated statistically by analysis of variance. The relative cyclin D1 levels were similar whether normalized to CD20 or cyclophilin A, indicating that CD20 levels are stable and can be used as a B-cell-specific normalizer. Statistically significant differences were found in the median levels of cyclin D1 mRNA (expressed as % CD20 mRNA) among cases of MCL (87.6), small lymphocytic lymphoma (9.9), follicular lymphoma (2.4), diffuse large B-cell lymphoma (5.9), marginal zone B-cell lymphoma (39.8), and Burkitt lymphoma (7.1) (P < 0.05). We conclude that qRT-PCR can be used to quantify cyclin D1 mRNA levels in archival tissue sections. Normalization of cyclin D1 to a B-cell-specific marker more accurately reflects overexpression by MCL than other methods that normalize using constitutively expressed mRNA species.
细胞周期蛋白D1过表达是诊断套细胞淋巴瘤（MCL）的一个重要标志物。我们采用实时定量逆转录聚合酶链反应（qRT-PCR）方法，使用ABI 7700 qRT-PCR系统，对手动显微切割的石蜡包埋组织切片中的细胞周期蛋白D1、CD20和亲环素A mRNA水平进行定量。研究组包括21例MCL和37例其他类型的B细胞非霍奇金淋巴瘤。将细胞周期蛋白D1 mRNA拷贝数标准化为CD20和亲环素A mRNA，并通过方差分析进行统计学评估。无论以CD20还是亲环素A作为标准化指标，细胞周期蛋白D1的相对水平相似，这表明CD20水平稳定，可作为B细胞特异性标准化指标。在MCL（87.6）、小淋巴细胞淋巴瘤（9.9）、滤泡性淋巴瘤（2.4）、弥漫性大B细胞淋巴瘤（5.9）、边缘区B细胞淋巴瘤（39.8）和伯基特淋巴瘤（7.1）病例中，细胞周期蛋白D1 mRNA的中位数水平（以% CD20 mRNA表示）存在统计学显著差异（P < 0.05）。我们得出结论，qRT-PCR可用于定量存档组织切片中的细胞周期蛋白D1 mRNA水平。与使用组成性表达的mRNA种类进行标准化的其他方法相比，将细胞周期蛋白D1标准化为B细胞特异性标志物能更准确地反映MCL中的过表达情况。
