Supplementary Material for: Proteomic Profiling of Diffuse Large B-Cell Lymphomas

<b><i>Objective:</i></b> The aim of this study was to identify differences in proteome profiles of diffuse large B-cell lymphoma (DLBCL) of nongerminal center (non-GC) versus GC type in the search for new markers and drug targets. <b><i>Methods:</i></b> Six DLBCL, with 3 repeats for each, were used for the initial study by proteomics: 3 non-GC and 3 GC DLBCL cases. For immunohistochemistry, tissue microarrays were made from 31 DLBCL samples: 16 non-GC de novo lymphomas and 15 GC cases (11 transformed from follicular lymphomas and 4 de novo GC lymphomas). Proteome profiling was performed by two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry. <b><i>Results:</i></b> Ninety-one proteins were found differentially expressed in non-GC compared to GC type. The Cytoscape tool was used for systemic analysis of proteomics data, revealing 19 subnetworks representing functions affected in non-GC versus GC types of DLBCL. <b><i>Conclusion:</i></b> A validation study of 3 selected proteins (BiP/Grp78, Hsp90, and cyclin B2) showed the enhanced expression in non-GC DLBCL, supporting the proteomics data.<b><i>研究目的：</i></b> 本研究旨在探究非生发中心（nongerminal center, non-GC）型与生发中心（germinal center, GC）型弥漫性大B细胞淋巴瘤（diffuse large B-cell lymphoma, DLBCL）的蛋白质组谱差异，以期发掘新型生物标志物与药物作用靶点。<b><i>方法：</i></b> 本研究的蛋白质组学初始实验纳入6例DLBCL样本，每例设置3次生物学重复，其中非GC型与GC型各3例。免疫组化实验采用31例DLBCL样本构建组织微阵列：其中16例为新发非GC型淋巴瘤，15例为GC型病例（含11例由滤泡性淋巴瘤转化而来的病例与4例新发GC型淋巴瘤）。蛋白质组谱分析采用二维凝胶电泳与基质辅助激光解吸电离飞行时间质谱（matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, MALDI-TOF MS）完成。<b><i>结果：</i></b> 相较于GC型DLBCL，非GC型样本中共鉴定出91个差异表达蛋白。借助Cytoscape工具对蛋白质组学数据进行系统分析，共识别出19个功能子网，用以表征非GC型与GC型DLBCL间存在功能异常的生物学过程。<b><i>结论：</i></b> 针对筛选出的3种蛋白（免疫球蛋白重链结合蛋白/葡萄糖调节蛋白78（BiP/Grp78）、热休克蛋白90（Hsp90）及细胞周期蛋白B2（cyclin B2））的验证实验结果显示，其在非GC型DLBCL中呈高表达，验证了蛋白质组学分析结果。