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利用“长距离”聚合酶链反应检测儿童伯基特淋巴瘤中的t(8;14)(q24;132)易位:一种诊断伯基特淋巴瘤的新方法

[Detection of translocation t(8;14)(q24;132) in pediatric Burkitt's lymphomas using "long distance" polymerase chain reaction: a new method for diagnosis of Burkitt's lymphomas].

作者信息

zur Stadt U, Reiter A, Welte K, Sykora K W

机构信息

Abteilung Pädiatrische Hämatologie und Onkologie, Medizinische Hochschule Hannover.

出版信息

Klin Padiatr. 1997 Jul-Aug;209(4):165-71. doi: 10.1055/s-2008-1043963.

DOI:10.1055/s-2008-1043963
PMID:9340426
Abstract

The pediatric Non-Hodgkin's lymphomas are a heterogeneous group of malignancies of B- or T-cell origin. Approximately half of them are characterized as Burkitt's lymphomas. Typically, one of the reciprocal translocations t(8;14)(q24;q32), t(2;8)(p11;q24) or t(8;22)(q24;q11) is seen in the tumor cell, each involving the protooncogene c-myc on chromosome 8. Characteristically, in most patients the translocation occurs between the distal end of the long arm on chromosome 8 (c-myc) and chromosome 14 (immunoglobulin heavy chain locus, IgH). The breakpoint regions are distributed over a wide range of more than 10 Kb on chr. 8 and over several hundred Kb on chr. 14. With standard-PCR, fragments can only be amplified to a size of about 2 Kb. The development of PCR-applications to generate long products up to 20 Kb now allows a detection of these breakpoints. Several primer pairs from different regions of the IgH-gene and the c-myc-gene were tested in each patient. Until now, 20 patients with Burkitt's Lymphoma or B-ALL characterized by L3 morphology were examined. All patients were treated according the protocols of the NHL-BFM '90 or '95 study. In 11/20 patients, recombinations between chromosomes 8 and 14 could be detected with our primer pairs. In serial dilutions of DNA from malignant cells in DNA from healthy controls, sensitivities of one malignant cell in 2 x 10(4) normal cells could be obtained. This method will now allow us to characterize the involved breakpoints more exactly and to analyze patient samples (blood, bone marrow, aphereses products and residual tumors) during or after therapy for the existence of minimal residual tumor cells.

摘要

儿童非霍奇金淋巴瘤是一组源于B细胞或T细胞的异质性恶性肿瘤。其中约一半被归类为伯基特淋巴瘤。通常,肿瘤细胞中可见相互易位之一t(8;14)(q24;q32)、t(2;8)(p11;q24)或t(8;22)(q24;q11),每种易位都涉及8号染色体上的原癌基因c-myc。其特征是,在大多数患者中,易位发生在8号染色体长臂远端(c-myc)与14号染色体(免疫球蛋白重链基因座,IgH)之间。断点区域分布在8号染色体上超过10kb的广泛范围内,在14号染色体上分布在数百kb范围内。使用标准PCR,片段只能扩增到约2kb大小。现在能够生成长达20kb长产物的PCR应用技术的发展,使得检测这些断点成为可能。在每位患者中测试了来自IgH基因和c-myc基因不同区域的几对引物。到目前为止,对20例具有L3形态特征的伯基特淋巴瘤或B-ALL患者进行了检查。所有患者均按照NHL-BFM '90或'95研究的方案进行治疗。在20例患者中的11例中,使用我们的引物对能够检测到8号和14号染色体之间的重组。在来自健康对照者DNA中的恶性细胞DNA系列稀释液中,可获得在2×10(4)个正常细胞中有1个恶性细胞的灵敏度。这种方法现在将使我们能够更准确地表征所涉及的断点,并在治疗期间或治疗后分析患者样本(血液、骨髓、采集产物和残留肿瘤)中是否存在微小残留肿瘤细胞。

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