Search for cancer markers from endometrial tissues using differentially labeled tags iTRAQ and clCAT with multidimensional liquid chromatography and tandem mass spectrometry
Search for cancer markers from endometrial tissues using differentially labeled tags iTRAQ and clCAT with multidimensional liquid chromatography and tandem mass spectrometry
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DOI：
10.1021/pr049821j
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发表时间：
2005-03-01
影响因子：
4.4
通讯作者：
Siu, KWM
中科院分区：
文献类型：
--
作者：
DeSouza, L;Diehl, G;Siu, KWM
利用多维液相色谱和串联质谱仪，结合差异标记标记iTRAQ和clCAT，从子宫内膜组织匀浆中发现并鉴定了9种潜在的子宫内膜癌标志物(EmCa)。在失活后15~20min内将组织在液氮中快速冷冻。进行蛋白质组学分析的样品在加工前用蛋白酶抑制剂处理。在EMCA中高表达的标志性蛋白有：伴侣蛋白10、丙酮酸激酶M1或M2同工酶、钙调素、异质性核糖核蛋白DO、巨噬细胞移动抑制因子和聚合性免疫球蛋白受体前体；低表达的有α-L抗胰蛋白酶前体、肌酸激酶B和转明胶。伴侣蛋白10的结果证实了我们之前使用表面增强激光解吸/电离质谱仪观察到的EmCa组织中的过度表达，并得到了Western分析和免疫组织化学的验证[Yang，E.C.C.et al.[J.蛋白质组研究2004，3,636-643]。用iTRAQ和clCAT标记观察到丙酮酸激酶过表达。所有九种标记物都被发现与各种形式的癌症有关。这些标记物和其他标记物的组合可以为诊断和筛查EMCA提供足够的选择性。使用clCAT导致鉴定出更高比例的低丰度信号蛋白；相反，iTRAQ导致更丰富的核糖体蛋白和转录因子的比例更高。
A total of nine potential markers for endometrial cancer (EmCa) have been discovered and identified from endometrial tissue homogenates using a combination of differentially labeled tags, iTRAQ and clCAT, with multidimensional liquid chromatography and tandem mass spectrometry. The tissues were snap frozen in liquid nitrogen within 15-20 min after devitalization. Samples for proteomic analysis were treated with protease inhibitors before processing. Marker proteins that were overexpressed in EmCa are chaperonin 10, pyruvate kinase M1 or M2 isozyme, calgizzarin, heterogeneous nuclear ribonucleoprotein DO, macrophage migratory inhibitory factor, and polymeric immunoglobulin receptor precursor; those that were underexpressed are alpha-l-antitrypsin precursor, creatine kinase B, and transgelin. The chaperonin 10 result confirms our earlier observation of overexpression in EmCa tissues using surface-enhanced laser desorption/ionization mass spectrometry, verified by Western analysis and immunohistochemistry [Yang, E. C. C. et al. J. Proteome Res. 2004, 3, 636-643]. Pyruvate kinase was observed to be overexpressed using both iTRAQ and clCAT labeling. All nine markers have been found to be associated with various forms of cancer. A panel of these plus other markers may confer sufficient selectivity for diagnosing and screening of EmCa. The use of clCAT led to identification of a higher proportion of lower-abundance signaling proteins; conversely, iTRAQ resulted in a higher percentage of the more abundant ribosomal proteins and transcription factors.