Proteomic analysis of hematopoietic stem cell-like fractions in leukemic disorders
Proteomic analysis of hematopoietic stem cell-like fractions in leukemic disorders
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DOI：
10.1038/sj.onc.1206855
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发表时间：
2003-08-28
期刊：
影响因子：
8
通讯作者：
Mano, H
中科院分区：
文献类型：
--
作者：
Ota, J;Yamashita, Y;Mano, H
DNA微阵列分析已被应用于人类血液系统恶性肿瘤的分子标记物的鉴定。然而，给定mRNA的丰度与编码蛋白质的丰度之间的相对低的相关性使得除了“转录组”之外，直接表征恶性细胞的蛋白质谱或“蛋白质组”是重要的。“为了鉴定白血病中特异性表达的蛋白质，我们从13名患有各种白血病疾病的个体的骨髓中分离出AC 133（+）造血干细胞样组分，并通过二维电泳比较它们的蛋白质谱。共检测到11个差异表达的蛋白质点，对应于10个独立的蛋白质，肽指纹图谱结合质谱分析这些蛋白质揭示它们包括NuMA（核蛋白，与有丝分裂器），热休克蛋白，和氧化还原调节剂。白血病原始细胞中NuMA的丰度与复杂核型异常的存在显著相关。NuMA在小鼠骨髓细胞系中的条件表达导致诱导非整倍体、细胞周期停滞在G（2）-M期和凋亡。这些结果表明，从新鲜的临床标本中获得的背景匹配的细胞组分的蛋白质组分析的潜力，以提供深入了解人类白血病发生的机制。
DNA microarray analysis has been applied to identify molecular markers of human hematological malignancies. However, the relatively low correlation between the abundance of a given mRNA and that of the encoded protein makes it important to characterize the protein profile directly, or 'proteome,' of malignant cells in addition to the 'transcriptome.' To identify proteins specifically expressed in leukemias, here we isolated AC133(+) hematopoietic stem cell-like fractions from the bone marrow of 13 individuals with various leukemic disorders, and compared their protein profiles by two-dimensional electrophoresis. A total of 11 differentially expressed protein spots corresponding to 10 independent proteins were detected, and peptide fingerprinting combined with mass spectrometry of these proteins revealed them to include NuMA (nuclear protein that associates with the mitotic apparatus), heat shock proteins, and redox regulators. The abundance of NuMA in the leukemic blasts was significantly related to the presence of complex karyotype anomalies. Conditional expression of NuMA in a mouse myeloid cell line resulted in the induction of aneuploidy, cell cycle arrest in G(2)-M phases, and apoptosis. These results demonstrate the potential of proteome analysis with background-matched cell fractions obtained from fresh clinical specimens to provide insight into the mechanism of human leukemogenesis.