儿童T淋巴母细胞淋巴瘤血清差异蛋白分析

Analysis of serum differential proteins in children with T-cell lymphoblastic lymphoma

目的应用基于同位素标记的相对和绝对定量技术(iTRAQ)筛选和鉴定儿童T淋巴母细胞淋巴瘤(T-LBL)与对照组的血清差异表达蛋白。方法收集T-LBL及健康儿童血清标本各20例,采用基于iTRAQ的蛋白质组学方法进行筛选、鉴定及生物信息学分析;对差异表达蛋白S100A8与富含亮氨酸α-2糖蛋白1(LRG1)应用ELISA进一步验证;采用受试者工作特征(ROC)曲线分析评价S100A8与LRG1的诊断效果。结果与正常儿童比较,T-LBL患儿血清中检测出73种差异表达倍数大于1.2倍的蛋白质(P<0.05),其中45种蛋白表达上调,28种蛋白表达下调;生物信息学分析显示这些差异蛋白主要参与组织损伤和异常、代谢疾病、心血管疾病、细胞间信号传导和交互作用、组织发育等相关的生物学功能;参与的主要信号通路包括急性期反应信号、LXR/RXR激活、动脉粥样硬化信号通路等。ELISA验证结果显示T-LBL血清S100A8与LRG1蛋白浓度高于正常对照,与蛋白质组学鉴定结果一致;ROC曲线分析结果显示,S100A8、LRG1、S100A8联合LRG1的ROC曲线下面积(AUC)分别为0.862、0.934、0.971。结论S100A8与LRG1是值得进一步研究的儿童T-LBL潜在生物标志物及治疗靶点。

Objective To screen and verify the serum differentially expressed proteins of children with T-lymphoblastic lymphoma(T-LBL)and control group by using isobaric tag for relative and absolute quantification(iTRAQ).Methods Serum protein expression profiles from 20 cases of normal healthy controls,and 20 cases of T-LBL patients were detected by iTRAQ labeling coupled with two-dimensional liquid chromatography–tandem mass spectrometry(2D LC–MS/MS),and analyzed by bioinformatics software.The differential expression of S100A8 and leucine rich repeatα-2 glycoprotein 1(LRG1)were validated by ELISA.Receiver operating characteristic(ROC)curve was used to evaluate the diagnostic effect of S100A8 and LRG1.Results Compared with normal healthy controls,73 differentially expressed(expressed more than 1.2 times,P<0.05)proteins in T-LBL serum were detected,of which 45 proteins were up-regulated and 28 proteins were down-regulated.Ingenuity Pathway Analysis(IPA)analysis revealed that these differentially expressed proteins were related to the function of Organismal Injury and Abnormalities、Metabolic Disease、Cardiovascular Disease、Cell-To-Cell Signaling and Interaction、Tissue Development etc.,and involved in a variety of signaling pathways,including acute phase response signaling、LXR/RXR activation、atherosclerosis signaling and so on.S100A8 and LRG1were found to be elevated in T-LBL patients when compared to the normal controls as examined by ELISA(P<0.05),which was consistent with the iTRAQ result.The overall predictive accuracy of each protein was reflected by the area under the ROC curve(AUC),S100A8、LRG1、and combinations of S100A8 and LRG1 had AUC of 0.862、0.934 and 0.971 respectively.Conclusions S100A8 and LRG1 may be serve as serum candidate biomarkers and therapeutic targets for pediatric T-LBL.