用于套细胞淋巴瘤诊断的血液、骨髓和组织标本中细胞周期蛋白D1信使核糖核酸的实时定量逆转录聚合酶链反应

Real-time quantitative reverse transcription-PCR for cyclin D1 mRNA in blood, marrow, and tissue specimens for diagnosis of mantle cell lymphoma.

作者信息

Howe John Greg, Crouch Jill, Cooper Dennis, Smith Brian R

机构信息

Department of Laboratory Medicine, Yale University School of Medicine, 333 Cedar St., New Haven, CT 06520, USA.

出版信息

Clin Chem. 2004 Jan;50(1):80-7. doi: 10.1373/clinchem.2003.024695. Epub 2003 Nov 21.

DOI:10.1373/clinchem.2003.024695

PMID:14633913

Abstract

BACKGROUND

Overexpression of cyclin D1 mRNA, found in mantle cell lymphoma (MCL), is a critical diagnostic marker. We investigated the use of real-time reverse transcription-PCR (RT-PCR) for cyclin D1.

METHODS

We studied 97 fresh specimens (50 blood, 30 bone marrow, 15 lymph node, and 2 other samples) from patients diagnosed with a variety of lymphoproliferative diseases, including 25 cases of MCL. We used real-time quantitative RT-PCR to evaluate cyclin D1 mRNA expression. Because blood and marrow specimens may contain only a minority of potentially malignant cells (as opposed to most lymph nodes) and to increase sensitivity, we normalized the cyclin D1 mRNA concentrations to mRNA of a B-cell-specific marker, CD19, as well as to previously characterized beta(2)-microglobulin mRNA.

RESULTS

In 16 of 21 cases of MCL with overt disease, the ratio of cyclin D1 mRNA to beta(2)-microglobulin mRNA was increased, but all 21 cases showed increased ratios of cyclin D1 mRNA to CD19 mRNA. Cyclin D1 mRNA was low or undetectable in various lymphoproliferative diseases, including cases of ambiguous immunophenotype. The mRNA ratios were stable over 3-7 days of sample storage.

CONCLUSION

Quantitative RT-PCR for cyclin D1 mRNA normalized to CD19 mRNA can be used in the diagnosis of MCL in blood, marrow, and tissue.

摘要

背景

在套细胞淋巴瘤（MCL）中发现的细胞周期蛋白D1信使核糖核酸（mRNA）过表达是一个关键的诊断标志物。我们研究了实时逆转录聚合酶链反应（RT-PCR）用于检测细胞周期蛋白D1的情况。

方法

我们研究了97份来自被诊断患有多种淋巴增殖性疾病患者的新鲜标本（50份血液、30份骨髓、15份淋巴结和2份其他样本），其中包括25例MCL。我们使用实时定量RT-PCR来评估细胞周期蛋白D1 mRNA的表达。由于血液和骨髓标本可能仅含有少数潜在恶性细胞（与大多数淋巴结相反），为了提高敏感性，我们将细胞周期蛋白D1 mRNA浓度相对于一种B细胞特异性标志物CD19的mRNA以及先前已鉴定的β2-微球蛋白mRNA进行标准化。

结果

在21例有明显疾病的MCL病例中，16例细胞周期蛋白D1 mRNA与β2-微球蛋白mRNA的比率升高，但所有21例细胞周期蛋白D1 mRNA与CD19 mRNA的比率均升高。在各种淋巴增殖性疾病中，包括免疫表型不明确的病例，细胞周期蛋白D1 mRNA水平较低或无法检测到。mRNA比率在样本储存3至7天内保持稳定。