Stoos-Veić Tajana, Livun Ana, Ajduković Radmila, Pejsa Vlatko, Jaksić Ozren, Kusec Rajko
Department of Cytology and Cytometry, Dubrava University Hospital, Zagreb, Croatia.
Coll Antropol. 2010 Jun;34(2):425-9.
According to WHO classification follicular lymphoma (FL) is a neoplasm composed of follicle centre (germinal centre) B-cells, which usually has at least a partially follicular pattern. Bone marrow (BM) infiltration by lymphoma occurs in 40-70% of cases at the time of diagnosis. The characteristic chromosomal translocation of follicular lymphoma is t(14;18)(q32;q21) with transposition of BCL2 oncogene to the regulatory region of immunoglobulin heavy chain gene IgH. Aim of this study was to determine the frequency of PCR detection of IgH/BCL2 in DNA samples isolated from archival cytological slides of lymph node aspirates, bone marrow and/or peripheral blood (PB) obtained from patients with histologically confirmed follicular lymphoma using primers and protocol proposed by BIOMED-2 consortium. We also compared molecular with cytomorphological findings in bone marrow/peripheral blood and tested this method of detection of IgH/BCL2 molecular marker in monitoring minimal residual disease (MRD) in routine clinical setting. DNA was successfully isolated from all archival cytological slides obtained by fine needle aspiration of lymph nodes as well as from 75% of smears of bone marrow aspirates from 19 patients. Fusion oncogene was detected in 10 of 19 patients (52%). For patients with PCR IgH/BCL2 positive lymph nodes, molecular test found BM infiltration in 5 cases (83%), while cytomorphology detected infiltration in three of eight cases (37%) available for comparison. May-Grünwald-Giemsa stained cytological smears can be used for PCR-based ancillary methods and the rate of detection of IgH/BCL2 rearrangement is similar to results reported for paraffin-embedded tissues. For patients with detectable baseline molecular marker, PCR is a highly suitable method for detection of bone marrow involvement and monitoring MRD.
根据世界卫生组织的分类，滤泡性淋巴瘤（FL）是一种由滤泡中心（生发中心）B细胞组成的肿瘤，通常至少具有部分滤泡样结构。在诊断时，40%-70%的病例存在淋巴瘤骨髓浸润。滤泡性淋巴瘤的特征性染色体易位是t(14;18)(q32;q21)，即BCL2癌基因转位至免疫球蛋白重链基因IgH的调控区。本研究的目的是使用BIOMED-2联盟提出的引物和方案，确定从组织学确诊为滤泡性淋巴瘤患者的淋巴结穿刺、骨髓和/或外周血（PB）存档细胞学载玻片分离的DNA样本中，IgH/BCL2的PCR检测频率。我们还比较了骨髓/外周血的分子学与细胞形态学结果，并在常规临床环境中测试了这种检测IgH/BCL2分子标志物的方法用于监测微小残留病（MRD）。通过细针穿刺淋巴结获得的所有存档细胞学载玻片以及19例患者75%的骨髓穿刺涂片均成功提取到DNA。19例患者中有10例（52%）检测到融合癌基因。对于PCR检测IgH/BCL2阳性的淋巴结患者，分子检测发现5例（83%）存在骨髓浸润，而细胞形态学在可供比较的8例中有3例（37%）检测到浸润。May-Grünwald-Giemsa染色的细胞学涂片可用于基于PCR的辅助方法，IgH/BCL2重排的检测率与石蜡包埋组织报道的结果相似。对于可检测到基线分子标志物的患者，PCR是检测骨髓受累和监测MRD的高度合适方法。
