Division of Hematology, Department of Internal Medicine, Aichi Medical University, Aichi, Japan.
Genes Chromosomes Cancer. 2012 Jan;51(1):42-53. doi: 10.1002/gcc.20929. Epub 2011 Oct 2.
In this study, we established and analyzed a novel human myeloid leukemia cell line, AMU-AML1, from a patient with acute myeloid leukemia with multilineage dysplasia before the initiation of chemotherapy. AMU-AML1 cells were positive for CD13, CD33, CD117, and HLA-DR by flow cytometry analysis and showed a single chromosomal abnormality, 46, XY, t(12;22)(p13;q11.2), by G-banding and spectral karyotyping. Fluorescent in situ hybridization analysis indicated that the chromosomal breakpoint in band 12p13 was in the sequence from the 5' untranslated region to intron 1 of TEL and that the chromosomal breakpoint in band 22q11 was in the 3' untranslated region of MN1. The chimeric transcript and protein of MN1-TEL could not be detected by reverse-transcriptase polymerase chain reaction or Western blot analysis. However, the MN1 gene was amplified to three copies detected by array comparative genomic hybridization analysis, and the expression levels of the MN1 transcript and protein were high in AMU-AML1 cells when compared with other cell lines with t(12;22)(p13;q11-12). Our data showed that AMU-AML1 cells contain t(12;22)(p13;q11.2) without chimeric fusion of MN1 and TEL. The AMU-AML1 cells gained MN1 copies and had high expression levels of MN1. Thus, the AMU-AML1 cell line is useful for studying the biological consequences of t(12;22)(p13;q11.2) lacking chimeric MN1-TEL.
在这项研究中,我们从一名在开始化疗前患有伴多系发育异常的急性髓系白血病的患者中建立并分析了一种新型的人髓系白血病细胞系 AMU-AML1。AMU-AML1 细胞通过流式细胞术分析呈 CD13、CD33、CD117 和 HLA-DR 阳性,并通过 G 带和光谱核型分析显示单一染色体异常,46,XY,t(12;22)(p13;q11.2)。荧光原位杂交分析表明,12p13 带中的染色体断裂点位于 TEL 的 5'非翻译区到内含子 1,22q11 带中的染色体断裂点位于 MN1 的 3'非翻译区。逆转录聚合酶链反应或 Western blot 分析未能检测到 MN1-TEL 的嵌合转录本和蛋白。然而,通过阵列比较基因组杂交分析检测到 MN1 基因扩增至三个拷贝,并且与具有 t(12;22)(p13;q11-12)的其他细胞系相比,AMU-AML1 细胞中的 MN1 转录本和蛋白表达水平较高。我们的数据表明,AMU-AML1 细胞含有 t(12;22)(p13;q11.2),但没有 MN1 和 TEL 的嵌合融合。AMU-AML1 细胞获得 MN1 拷贝,并具有高水平的 MN1 表达。因此,AMU-AML1 细胞系可用于研究缺乏嵌合 MN1-TEL 的 t(12;22)(p13;q11.2)的生物学后果。