Taniwaki M, Sliverman G A, Nishida K, Horiike S, Misawa S, Shimazaki C, Miura I, Nagai M, Abe M, Fukuhara S
Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Japan.
Blood. 1995 Aug 15;86(4):1481-6.
Translocation of the BCL2 gene in B-cell malignancies carrying t(14;18) and amplification of the BCL2 gene in a cell line (HBL-2) derived from a non-Hodgkin's lymphoma (NHL) were detected specifically in both metaphase spreads and interphase nuclei by fluorescence in situ hybridization (FISH) using yeast artificial chromosomes (YACs). A YAC clone containing the BCL2 gene yA153A6, a 360-kb clone spanning from approximately 60 kb upstream of BCL2 exon 1 to approximately 60 kb 3' of the minor breakpoint cluster region, was used for single-color FISH analysis. Seven patients with NHL and one patient with acute lymphoblastic leukemia were analyzed for BCL2 translocations. Interphase nuclei of NHL patients showed three signals when hybridized with the yA153A6 probe. This was expected because the YAC clone spans the BCL2 breakpoint regions on 18q21.3. In a patient with acute lymphoblastic leukemia, a positive signal for BCL2 was detected on der(14) at band 14q32.33 by single-color FISH with the yA153A6 probe, whereas no signals were detected on der(18). The amplification of BCL2 in the HBL-2 cell line was observed on a characteristic abnormal chromosome 18, add(18)(q23); the periodic pattern of the fluorescent signal of this region was suggestive of an amplicon. Using double-color FISH with YAC clones containing the more centromeric 18q21.3 gene gastrin-releasing peptide (y302F10) and the 14q32.33 gene (IgH; Y6), we detected t(14;18) by showing the juxtaposition of the 18q21.3 and 14q32.33 bands on the derivative chromosome 18. Interphase FISH with these YAC clones provided a rapid procedure for the diagnosis of B-cell malignancies carrying t(14;18). In addition, we showed that translocations and amplification of the BCL2 gene can be detected at the single-cell level.
利用酵母人工染色体（YAC）通过荧光原位杂交（FISH）技术，在中期染色体铺展和间期细胞核中均特异性检测到了携带t(14;18)的B细胞恶性肿瘤中BCL2基因的易位，以及源自非霍奇金淋巴瘤（NHL）的细胞系（HBL-2）中BCL2基因的扩增。一个包含BCL2基因yA153A6的YAC克隆，这是一个360 kb的克隆，从BCL2外显子1上游约60 kb延伸至微小断点簇区域下游约60 kb，用于单色FISH分析。对7例NHL患者和1例急性淋巴细胞白血病患者进行了BCL2易位分析。NHL患者的间期细胞核与yA153A6探针杂交时显示出三个信号。这是预期的结果，因为YAC克隆跨越了18q21.3上的BCL2断点区域。在1例急性淋巴细胞白血病患者中，通过用yA153A6探针进行单色FISH，在der(14)的14q32.33带检测到BCL2的阳性信号，而在der(18)上未检测到信号。在HBL-2细胞系中，在一条特征性异常染色体18，add(18)(q23)上观察到BCL2的扩增；该区域荧光信号的周期性模式提示存在扩增子。使用包含更靠近着丝粒的18q21.3基因胃泌素释放肽（y302F10）和14q32.33基因（IgH；Y6）的YAC克隆进行双色FISH，我们通过显示衍生染色体18上18q21.3和14q32.33带的并列检测到了t(14;18)。用这些YAC克隆进行间期FISH为携带t(14;18)的B细胞恶性肿瘤的诊断提供了一种快速方法。此外，我们还表明，BCL2基因的易位和扩增可以在单细胞水平上检测到。
