Medical Proteome-Center, University of Bochum, Bochum, Germany; Department of Internal Medicine, IMBL, Knappschaftskrankenhaus, University of Bochum, Bochum, Germany.
Proteomics Clin Appl. 2007 Jan;1(1):47-61. doi: 10.1002/prca.200600491. Epub 2006 Dec 5.
Improved methods for the early diagnosis of colorectal cancer by way of sensitive and specific tumour markers are highly desirable. Therefore, efficient strategies for biomarker discovery are urgently needed. Here we present an approach that is based on the direct experimental access to proteins released by SW620 human colorectal cancer cells in vitro. A 2-D map and a catalogue of this subproteome - here termed the secretome - were established comprising more than 320 identified proteins which translate into approximately 220 distinct genes. As the majority of the secretome constituents were nominally cellular proteins, we directly compared the secretome and the total proteome by 2-D-DIGE analysis. We provide evidence that unspecific release through cell death, classical secretion, ectodomain shedding, and exosomal release contribute to the secretome in vitro, presumably reflecting the mechanisms in vivo which lead to the occurrence of tumour-specific proteins in the circulation. These data together with the fact that the SW620 secretome catalogue, as presented here, does comprise a large number of known and novel biomarker candidates, validates our approach to isolate and characterize the tumour cell secretome in vitro as a rich source for tumour biomarkers.
人们迫切需要通过敏感和特异的肿瘤标志物来改善结直肠癌的早期诊断方法。因此,高效的生物标志物发现策略迫在眉睫。在这里,我们提出了一种基于直接实验获取人结直肠癌细胞 SW620 体外释放蛋白的方法。建立了一个包含 320 多种已鉴定蛋白的 2-D 图谱和一个亚蛋白质组目录,这些蛋白可翻译成大约 220 种不同的基因。由于分泌组的大多数成分是名义上的细胞蛋白,我们通过 2-D-DIGE 分析直接比较了分泌组和总蛋白质组。我们提供的证据表明,通过细胞死亡、经典分泌、胞外域脱落和外泌体释放的非特异性释放,可能反映了体内导致肿瘤特异性蛋白在循环中出现的机制,导致体外分泌组中存在。这些数据以及 SW620 分泌组目录(如本文所述)确实包含大量已知和新的生物标志物候选物的事实,验证了我们分离和鉴定体外肿瘤细胞分泌组作为肿瘤生物标志物丰富来源的方法。