Integrated Proteomic Profiling of Cell Line Conditioned Media and Pancreatic Juice for the Identification of Pancreatic Cancer Biomarkers
Integrated Proteomic Profiling of Cell Line Conditioned Media and Pancreatic Juice for the Identification of Pancreatic Cancer Biomarkers
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DOI：
10.1074/mcp.m111.008599
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发表时间：
2011-10-01
影响因子：
7
通讯作者：
Diamandis, Eleftherios P.
中科院分区：
文献类型：
--
作者：
Makawita, Shalini;Smith, Chris;Diamandis, Eleftherios P.
胰腺癌是癌症相关死亡的主要原因之一，迫切需要血清学生物标志物。大多数发现阶段的研究侧重于使用一种生物来源进行分析。本研究详细介绍了胰腺癌相关细胞系条件培养基和胰液的联合挖掘，以确定推定的诊断线索。使用强阳离子交换色谱，然后在LTQ-Orbitrap质谱仪上进行LC-MS/MS，我们广泛地表征了来自六种胰腺癌细胞系（BxPc 3、MIA-PaCa 2、PANC 1、CAPAN 1、CFPAC 1和SU. 86. 86）、正常人胰腺导管上皮细胞系HPDE和来自导管腺癌患者的六种胰液样品的两个池的条件培养基的蛋白质组。所有样品均一式三份进行分析。在每个细胞系中，用两种或更多种肽鉴定了1261至2171种蛋白质，并且在胰液池中鉴定了平均521种蛋白质。总的来说，3479非冗余蛋白质被确定为高置信度，其中40%是细胞外或细胞膜结合的基因组本体分类的基础上。采用三种策略来鉴定候选生物标志物：（1）使用无标记蛋白质定量来检查癌症细胞系与正常细胞系之间的差异蛋白质表达，（2）综合分析，集中于多种生物流体之间的蛋白质重叠，以及（3）通过挖掘可药用数据库来进行组织特异性分析。使用ELISA对来自胰腺癌患者和健康对照的血浆样品中的前梯度同源物2、syncollin、嗅介蛋白-4、多聚免疫球蛋白受体和胶原α-l（VI）链的初步验证显示，来自胰腺癌患者的血浆中这些蛋白质显著增加（p < 0.01）。这五种蛋白质的组合显示出与单独的CA 19.9相比的受试者工作特征曲线下面积的改善。这些蛋白质的进一步验证是必要的，因为是其余组的候选人的调查。Molecular & Cellular Proteomics 10：10.1074/mcp. M111.008599，1-20，2011.
Pancreatic cancer is one of the leading causes of cancer-related deaths, for which serological biomarkers are urgently needed. Most discovery-phase studies focus on the use of one biological source for analysis. The present study details the combined mining of pancreatic cancer-related cell line conditioned media and pancreatic juice for identification of putative diagnostic leads. Using strong cation exchange chromatography, followed by LC-MS/MS on an LTQ-Orbitrap mass spectrometer, we extensively characterized the proteomes of conditioned media from six pancreatic cancer cell lines (BxPc3, MIA-PaCa2, PANC1, CAPAN1, CFPAC1, and SU.86.86), the normal human pancreatic ductal epithelial cell line HPDE, and two pools of six pancreatic juice samples from ductal adenocarcinoma patients. All samples were analyzed in triplicate. Between 1261 and 2171 proteins were identified with two or more peptides in each of the cell lines, and an average of 521 proteins were identified in the pancreatic juice pools. In total, 3479 nonredundant proteins were identified with high confidence, of which similar to 40% were extracellular or cell membrane-bound based on Genome Ontology classifications. Three strategies were employed for identification of candidate biomarkers: (1) examination of differential protein expression between the cancer and normal cell lines using label-free protein quantification, (2) integrative analysis, focusing on the overlap of proteins among the multiple biological fluids, and (3) tissue specificity analysis through mining of publically available databases. Preliminary verification of anterior gradient homolog 2, syncollin, olfactomedin-4, polymeric immunoglobulin receptor, and collagen alpha-1(VI) chain in plasma samples from pancreatic cancer patients and healthy controls using ELISA, showed a significant increase (p < 0.01) of these proteins in plasma from pancreatic cancer patients. The combination of these five proteins showed an improved area under the receiver operating characteristic curve to CA19.9 alone. Further validation of these proteins is warranted, as is the investigation of the remaining group of candidates. Molecular & Cellular Proteomics 10: 10.1074/mcp.M111.008599, 1-20, 2011.