基因组DNA的长距离扩增可检测间变性大细胞淋巴瘤中的t(2;5)(p23;q35)，但在其他非霍奇金淋巴瘤、霍奇金病或淋巴瘤样丘疹病中则无法检测到。

Long-range amplification of genomic DNA detects the t(2;5)(p23;q35) in anaplastic large-cell lymphoma, but not in other non-Hodgkin's lymphomas, Hodgkin's disease, or lymphomatoid papulosis.

作者信息

Sarris A H, Luthra R, Papadimitracopoulou V, Waasdorp M, Dimopoulos M A, McBride J A, Cabanillas F, Duvic M, Deisseroth A, Morris S W, Pugh W C

机构信息

Department of Hematology, University of Texas M. D. Anderson Cancer Center, Houston, USA.

出版信息

Ann Oncol. 1997;8 Suppl 2:59-63.

PMID:9209643

Abstract

DESIGN

Determine the frequency of t(2;5)(p23;q35) in anaplastic large-cell lymphoma (ALCL), non-Hodgkin's lymphoma (NHL), Hodgkin's disease (HD), and lymphomatoid papulosis (LP).

PATIENTS AND METHODS

The t(2;5) was detected with a long-range nested polymerase chain reaction (PCR) using 0.5 microgram of DNA (60,000-80,000 cells), 5'-primers from the NPM gene, 3'-primers from the ALK gene, agarose electrophoresis, hybridization, and autoradiography. Patients were evaluable if a 3016 base pair amplicon could be generated from tumor DNA with beta-globin primers.

RESULTS

Amplicons were detected by PCR of genomic DNA from three ALCL cell lines and four primary ALCLs known to t(2;5) positive. DNA from t(2;5)-positive cell lines diluted 10(4)-fold or 10(5)-fold generated amplicons in 100% or 20% of reactions, respectively. Archival tumor DNA from 144 patients was amplifiable by beta-globin amplicons in 126 (88%) who are considered evaluable for this study. Twenty-two had ALCL, 69 other NHLs, 30 HD, and five LP. Genomic DNA PCR detected the t(2;5) in 5 of 22 with ALCL (23%, 95% confidence intervals [95% CI] 8%-45%) but not in those with NHLs, HD, or LP. Among ALCLs the t(2;5) was confined to 5 of 20 with nodal presentations (25%, 95% CI 9%-49%), among whom it was seen in 5 of 15 with T-cell or null-cell phenotype (33%, 95% CI 12%-62%), in 4 of 11 with age < 40 years (36%, 95% CI 11%-69%), and in 4 of 9 with nodal presentations, T-cell or null-cell phenotype, and age < 40 years (44%, 95% CI 14%-79%). Amplicon sizes were different between cell lines and patients, reflecting unique genomic DNA breakpoints, as confirmed by DNA sequencing, and served as an internal control against specimen cross-contamination in the laboratory.

CONCLUSIONS

Long-range PCR of genomic DNA detects t(2;5) only in ALCL, but not in other NHLs, HD, or LP. Long-range PCR may be useful in establishing diagnosis, determining prognosis, and monitoring minimal residual disease in ALCL.

摘要

设计

确定间变性大细胞淋巴瘤（ALCL）、非霍奇金淋巴瘤（NHL）、霍奇金病（HD）和淋巴瘤样丘疹病（LP）中t(2;5)(p23;q35)的频率。

患者和方法

使用0.5微克DNA（60,000 - 80,000个细胞）、来自NPM基因的5'引物、来自ALK基因的3'引物、琼脂糖电泳、杂交和放射自显影，通过长程巢式聚合酶链反应（PCR）检测t(2;5)。如果用β-珠蛋白引物能从肿瘤DNA产生3016碱基对的扩增子，则患者可进行评估。

结果

通过PCR检测到来自三个已知t(2;5)阳性的ALCL细胞系和四个原发性ALCL的基因组DNA的扩增子。t(2;5)阳性细胞系的DNA稀释10⁴倍或10⁵倍后，分别在100%或20%的反应中产生扩增子。144例患者的存档肿瘤DNA中，126例（88%）可通过β-珠蛋白扩增子扩增，这些患者被认为可用于本研究评估。其中22例为ALCL，69例为其他NHL，30例为HD，5例为LP。基因组DNA PCR在22例ALCL中的5例中检测到t(2;5)（23%，95%置信区间[95%CI]8% - 45%），但在NHL、HD或LP患者中未检测到。在ALCL中，t(2;5)仅限于20例有淋巴结表现的患者中的5例（25%，95%CI 9% - 49%），其中15例T细胞或无细胞表型的患者中有5例（33%，95%CI 12% - 62%），11例年龄<40岁的患者中有4例（36%，95%CI 11% - 69%），9例有淋巴结表现、T细胞或无细胞表型且年龄<40岁的患者中有4例（44%，95%CI 14% - 79%）。细胞系和患者之间的扩增子大小不同，反映了独特的基因组DNA断点，经DNA测序证实，并作为实验室标本交叉污染的内部对照。