Taniguchi T, Fujita A, Takahashi S, Uchimaru K, Yoshikawa M, Asano S, Fujita T, Motokura T
Fourth Department of Internal Medicine, University of Tokyo, School of Medicine.
Jpn J Cancer Res. 1998 Feb;89(2):159-66. doi: 10.1111/j.1349-7006.1998.tb00544.x.
t(11;14)(q13;q32) observed in B-cell malignancies is associated with cyclin D1 (bcl-1, PRAD1, CCND1) overexpression. We devised a simple competitive reverse transcription-polymerase chain reaction (RT-PCR) assay for rapid detection of cyclin D1 overexpression. Sharing a single upstream primer derived from a homologous sequence in cyclins D1, D2 and D3, each PCR product serves as a competitor and cyclin D1 overexpression is determined by comparing the intensities of the three amplified products. We analyzed cyclin D1 in clinical specimens from 104 patients with lymphoid malignancies. Cyclin D1 overexpression was evident in 13 of 104 (7/72 non-Hodgkin's lymphomas, 0/6 adult T-cell lymphoma/leukemias, 0/4 Hodgkin's diseases, 0/11 acute lymphoblastic leukemias, 3/4 multiple myelomas, 1/2 Waldenström's macroglobulinemias, 1/2 prolymphocytic leukemias and 1/3 chronic lymphocytic leukemias). Among 72 patients for whom cytogenetic studies had been done, all 7 patients with t(11;14) were positive. The relative expression levels of D-type cyclins altered dramatically in the presence of t(11;14). Thus, this RT-PCR assay can identify tumors with cyclin D1 overexpression. Cyclin D1 overexpression was frequent in extranodal specimens (11 out of 32 vs. 2 of 72 lymph nodes) and was restricted to specific types of lymphoid malignancies, as observed using other methods. This reliable assay should be suitable to provide clinical guidance for the diagnosis and management of lymphoid malignancies, especially in the case of extranodal involvement.
在B细胞恶性肿瘤中观察到的t(11;14)(q13;q32)与细胞周期蛋白D1（bcl-1、PRAD1、CCND1）过表达相关。我们设计了一种简单的竞争性逆转录-聚合酶链反应（RT-PCR）检测方法，用于快速检测细胞周期蛋白D1过表达。每个PCR产物共享一个源自细胞周期蛋白D1、D2和D3同源序列的单一上游引物，作为竞争者，通过比较三种扩增产物的强度来确定细胞周期蛋白D1过表达。我们分析了104例淋巴系统恶性肿瘤患者临床标本中的细胞周期蛋白D1。104例中有13例出现细胞周期蛋白D1过表达（72例非霍奇金淋巴瘤中的7例、6例成人T细胞淋巴瘤/白血病中的0例、4例霍奇金病中的0例、11例急性淋巴细胞白血病中的0例、4例多发性骨髓瘤中的3例、2例华氏巨球蛋白血症中的1例、2例原淋巴细胞白血病中的1例和3例慢性淋巴细胞白血病中的1例）。在72例进行了细胞遗传学研究的患者中，所有7例t(11;14)患者均为阳性。在存在t(11;14)的情况下，D型细胞周期蛋白的相对表达水平发生了显著变化。因此，这种RT-PCR检测方法可以识别细胞周期蛋白D1过表达的肿瘤。如使用其他方法所观察到的，细胞周期蛋白D1过表达在结外标本中很常见（32例中的11例与72例淋巴结中的2例），并且仅限于特定类型的淋巴系统恶性肿瘤。这种可靠的检测方法应适合为淋巴系统恶性肿瘤的诊断和管理提供临床指导，尤其是在结外受累的情况下。
