Biomarker discovery from pancreatic cancer secretome using a differential proteomic approach
Biomarker discovery from pancreatic cancer secretome using a differential proteomic approach
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DOI：
10.1074/mcp.m500178-mcp200
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发表时间：
2006-01-01
影响因子：
7
通讯作者：
Pandey, A
中科院分区：
文献类型：
--
作者：
Gronborg, M;Kristiansen, TZ;Pandey, A
定量蛋白质组学可以作为一种筛选工具，用于鉴定作为癌症潜在生物标志物的差异表达蛋白。这些研究的候选生物标志物随后可以使用其他技术进行测试，用于癌症的早期检测。在这里，我们展示了使用细胞培养中氨基酸的稳定同位素标记（SILAC）方法来比较来自胰腺癌细胞的分泌蛋白（分泌组）与来自非肿瘤胰腺导管细胞的分泌蛋白（分泌组）。我们鉴定了145种差异分泌蛋白（> 1.5倍变化），其中一些先前被报道为胰腺癌中上调（如组织蛋白酶D，巨噬细胞集落刺激因子和纤维连接蛋白受体）或下调（如profilin 1和IGFBP-7）蛋白，证实了我们方法的有效性。此外，我们还发现了几种先前未与胰腺癌相关的蛋白，包括perlecan （HSPG2）、CD9抗原、纤维连接蛋白受体（整合素β 1）和一种被指定为预测成骨细胞蛋白（FAM3C）的新型细胞因子。这些新蛋白子集的差异表达通过Western blot分析得到验证。此外，使用胰腺癌组织微阵列进行免疫组化标记，证实了几种先前未被描述为在人类胰腺癌中升高的蛋白（CD9、perlecan、SDF4、apoE和纤维连接蛋白受体）的过表达，这表明这些蛋白可以进一步作为潜在的生物标志物进行研究。最后，将SILAC的蛋白表达数据与基因表达芯片获得的两种细胞系（Panc1和人胰管上皮细胞）的mRNA表达数据进行比较，得出相关系数(r)为0.28，证实了之前报道的RNA和蛋白表达研究之间的低相关性。
Quantitative proteomics can be used as a screening tool for identification of differentially expressed proteins as potential biomarkers for cancers. Candidate biomarkers from such studies can subsequently be tested using other techniques for use in early detection of cancers. Here we demonstrate the use of stable isotope labeling with amino acids in cell culture (SILAC) method to compare the secreted proteins (secretome) from pancreatic cancer-derived cells with that from non-neoplastic pancreatic ductal cells. We identified 145 differentially secreted proteins (> 1.5-fold change), several of which were previously reported as either up-regulated ( e. g. cathepsin D, macrophage colony stimulation factor, and fibronectin receptor) or down-regulated ( e. g. profilin 1 and IGFBP-7) proteins in pancreatic cancer, confirming the validity of our approach. In addition, we identified several proteins that have not been correlated previously with pancreatic cancer including perlecan (HSPG2), CD9 antigen, fibronectin receptor ( integrin beta 1), and a novel cytokine designated as predicted osteoblast protein (FAM3C). The differential expression of a subset of these novel proteins was validated by Western blot analysis. In addition, overexpression of several proteins not described previously to be elevated in human pancreatic cancer ( CD9, perlecan, SDF4, apoE, and fibronectin receptor) was confirmed by immunohistochemical labeling using pancreatic cancer tissue microarrays suggesting that these could be further pursued as potential biomarkers. Lastly the protein expression data from SILAC were compared with mRNA expression data obtained using gene expression microarrays for the two cell lines ( Panc1 and human pancreatic duct epithelial), and a correlation coefficient (r) of 0.28 was obtained, confirming previously reported poor associations between RNA and protein expression studies.