Guo Ye, Fan Bao-Li, Chen Yu-Mei, Hu Ying, Zou Yao, Chen Xiao-Juan, Zhang Li, Zhu Xiao-Fan
Diagnostic and Therapeutic Center for Childhoood Hematologic Diseases, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjing 300020, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Feb;18(1):116-21.
The study was aimed to establish differential proteomic expression analysis of two dimensional electrophoresis and mass spectrography, and to further explore the mechanisms of nosogenesis in childhood TEL/AML1(+) acute lymphoblastic leukemia. On the basis of initial leukocyte count and prognostic factors, patients enrolled in this study were divided into three risk groups (early relapse, high leukocyte count and standard risk groups). The proteins of leukemic cells from patients at diagnosis were separated by two-dimensional gel electrophoresis. Spot detection, quantification and alignment were performed with the PDQuest 7.3.0 image analysis software. Differentially expressed spots were analyzed by mass spectrometry for peptide mass finger (PMF) data. The results showed that the significant difference of protein expression profile existed in 3 groups of childhood TEL/AML1-positive acute lymphoblastic leukemia. Compared with the high leukocyte count and standard risk groups, 71 protein spots disappeared; 93 new protein spots were found; 37 protein spot expressions were up-regulated; 23 protein spots were down-regulated in early relapse group. Compared the high leukocyte count group, 6 protein spots disappeared, 56 new protein spots were found, 7 protein spot expression were up-regulated, 19 protein spot expressions were down-regulated in standard risk group. The identification of 40 differential protein spots by using mass spectrometry revealed some proteins in 3 groups such as tropomyosin, lactotransferrin, lactate dehydrogenase A, CRMP1 protein, alphaenolase, AKR1B1, calnexin precursor, heat shock protein (HSP86/HSP89/HSP90), annexin VI, G22P1 and so on. Among them the HSP86/HSP89/HSP90 highly expressed only in early reapse group, the lactotransferrin, alphenolase and G22P1 expressions were up-regulated in early relapse group. It is concluded that the protein expression in early relapse group is significant different from the others. Some proteins may be further used in research on leukemia mechanisms. In addition, these analyses may promote the identification of new targets for individualized treatment approaches.
本研究旨在建立二维电泳和质谱的差异蛋白质组表达分析方法，并进一步探讨儿童TEL/AML1(+)急性淋巴细胞白血病的发病机制。根据初始白细胞计数和预后因素，将本研究纳入的患者分为三个风险组（早期复发组、高白细胞计数组和标准风险组）。采用二维凝胶电泳分离诊断时患者白血病细胞的蛋白质。使用PDQuest 7.3.0图像分析软件进行斑点检测、定量和比对。对差异表达的斑点进行质谱分析以获取肽质量指纹（PMF）数据。结果显示，儿童TEL/AML1阳性急性淋巴细胞白血病3组间蛋白质表达谱存在显著差异。与高白细胞计数组和标准风险组相比，早期复发组有71个蛋白质斑点消失；发现93个新的蛋白质斑点；37个蛋白质斑点表达上调；23个蛋白质斑点表达下调。与高白细胞计数组相比，标准风险组有6个蛋白质斑点消失，发现56个新的蛋白质斑点，7个蛋白质斑点表达上调，19个蛋白质斑点表达下调。通过质谱鉴定40个差异蛋白质斑点，发现3组中存在一些蛋白质，如原肌球蛋白、乳铁传递蛋白、乳酸脱氢酶A、CRMP1蛋白、α烯醇化酶、AKR1B1、钙连接蛋白前体、热休克蛋白（HSP86/HSP89/HSP90）、膜联蛋白VI、G22P1等。其中HSP86/HSP89/HSP90仅在早期复发组高表达，乳铁传递蛋白、α烯醇化酶和G22P1在早期复发组表达上调。结论是早期复发组的蛋白质表达与其他组有显著差异。一些蛋白质可能进一步用于白血病机制的研究。此外，这些分析可能有助于识别个体化治疗方法的新靶点。
