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通过逆转录聚合酶链反应检测B细胞非霍奇金淋巴瘤中免疫球蛋白重链和BCL6基因的嵌合转录本。

Detection of chimaeric transcripts of the immunoglobulin heavy chain and BCL6 genes by reverse-transcriptase polymerase chain reaction in B-cell non-Hodgkin's lymphomas.

作者信息

Kawamata N, Nakamura Y, Miki T, Sato E, Isobe Y, Furusawa S, Hirosawa S, Oshimi K

机构信息

Department of Medicine, Juntendo University, School of Medicne, Tokyo, Japan.

出版信息

Br J Haematol. 1998 Mar;100(3):484-9. doi: 10.1046/j.1365-2141.1998.00603.x.

Abstract

T(3;14)(q27;q32) is frequently detected in B-cell non-Hodgkin's lymphomas, especially the diffuse large cell type and the follicular type. The BCL6 gene encoding a putative transcriptional factor which resides on 3q27 rearranges to the immunoglobulin heavy chain (IgH) gene on 14q32 in this chromosomal translocation. The upstream regulatory region of the BCL6 gene is replaced by the IgH gene. Deregulation of the BCL6 gene may contribute to tumourigenesis of these diseases. The rearrangement between the IgH and BCL6 genes generates chimaeric transcripts in which the joining (J) region of the IgH gene fuses to exon 3 of the BCL6 gene. We established a method to detect these chimaeric transcripts by reverse transcriptase polymerase chain reaction (RT-PCR) using the consensus sequence of the J region and the sequence of exon 3 of the BCL6 gene as primers. Using the semi-nested RT-PCR method and a cell line carrying t(3;14)(q27;q32), we detected one lymphoma cell among 10,000 background cells. We detected these chimaeric transcripts in two out of 13 clinical samples by this method. This method can detect t(3;14)(q27;q32) easily, whereas this alteration is frequently overlooked by routine karyotype analysis. Since this technique is sensitive enough to detect a small number of lymphoma cells with this genetic abnormality, it could be employed to detect contaminating lymphoma cells in bone marrow and peripheral blood and minimal residual diseases.

摘要

T(3;14)(q27;q32)在B细胞非霍奇金淋巴瘤中经常被检测到,尤其是弥漫大细胞型和滤泡型。在这种染色体易位中,位于3q27的编码假定转录因子的BCL6基因重排至14q32的免疫球蛋白重链(IgH)基因。BCL6基因的上游调控区域被IgH基因取代。BCL6基因的失调可能促成这些疾病的肿瘤发生。IgH和BCL6基因之间的重排产生嵌合转录本,其中IgH基因的连接(J)区域与BCL6基因的外显子3融合。我们建立了一种方法,使用J区域的共有序列和BCL6基因外显子3的序列作为引物,通过逆转录聚合酶链反应(RT-PCR)检测这些嵌合转录本。使用半巢式RT-PCR方法和携带t(3;14)(q27;q32)的细胞系,我们在10000个背景细胞中检测到一个淋巴瘤细胞。通过这种方法,我们在13个临床样本中的2个中检测到了这些嵌合转录本。这种方法可以轻松检测t(3;14)(q27;q32),而这种改变在常规核型分析中经常被忽视。由于该技术足够灵敏,能够检测到少量具有这种基因异常的淋巴瘤细胞,因此可用于检测骨髓和外周血中的污染淋巴瘤细胞以及微小残留病。

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