同位素标记联合二维液相色谱串联质谱技术筛选T淋巴细胞急性白血病特异标记物

ITRAQ Coupled with 2DLC-MS/MS to Screen and Identify Specific Bio-markers of T-Cell Acute Lymphoblastic Leukemia

目的:筛选并鉴定T淋巴细胞白血病(T-ALL)的可能标记物。方法:分别收集并纯化初治儿童B淋巴细胞白血病(B-ALL)与T-ALL血清各20份。应用同位素标记联合二维液相色谱串联质谱(i TRAQ-2DLC-MS/MS)和生物信息分析软件(IPA)筛查T-ALL特异性分子标志物。采用ELISA双抗夹心法分别检测上述40例样本中血清可溶性L-选择素(s L-selectin)表达水平,进一步验证2DLC-MS/MS技术的准确性,并计算其统计学意义。结果:有定量信息的非冗余蛋白468个,T-ALL组血清差异蛋白共有38个,其中显著上调蛋白质31个,下调蛋白质7个;T-ALL中s L-selectin表达明显上调,并得到ELISA检测结果证实。结论:T-ALL与B-ALL比较存在相对特异的血清差异蛋白,s L-selectin可作为T-ALL新的肿瘤分子标志物及潜在的治疗靶点。

Objective： To screen and identify potential biomarkers specific for T-cell acute lymphoblastic leukemia（ T-ALL）. Methods： Sera were collected from 20 newly diagnosed B-cell acute lymphoblastic leukemia（ B-ALL） patients and 20 T-ALL patients. Proteins were extracted,purified and digested with trypsin. All specimens were analyzed by isobaric tags for relative and absolute quantification（ i TRAQ） and two-dimensional liquid chromatography-tandem mass spectrometry（ 2 DLC-MS/MS） in a data-dependent mode. Enzyme-linked immunosorbent assay（ ELISA） was used to analyze the expression of serum soluble L-selectin（ sL-selectin）. Results： A total of 468 proteins were identified from distinct peptides. Compared with B-ALL group,31 proteins were significantly differentially up-regulated while 7 proteins were significantly down-regulated in T-ALL group,s L-selectin was the higher up-regulated in these differential expression proteins. The overexpression of s L-selectin in T-ALL was verified by ELISA. Conclusion： There are the differentially expressed proteins between T-ALL and B-ALL,and the s L-selectin is specific for T-ALL,which can not only become a newbiomarker for the diagnosis and prognosis of T-ALL,but also can be used as a potential target for therapy of this leukemia.