Secretome protein signature of human pancreatic cancer stem-like cells
Secretome protein signature of human pancreatic cancer stem-like cells
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DOI：
10.1016/j.jprot.2016.01.017
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发表时间：
2016-05-16
影响因子：
3.3
通讯作者：
Cecconi, Daniela
中科院分区：
文献类型：
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作者：
Brandi, Jessica;Dalla Pozza, Elisa;Cecconi, Daniela
新兴研究表明，胰腺导管腺癌 (PDAC) 含有癌症干细胞 (CSC) 亚群，其特征是自我更新、不依赖贴壁生长、长期增殖和化疗耐药。胰腺 CSC 的分泌蛋白组分析尚未进行，尽管它可以提供对肿瘤/微环境相互作用和细胞内过程的深入了解，并识别潜在的生物标志物。为了表征胰腺 CSC 的分泌蛋白，我们进行了基于 iTRAQ 的蛋白质组分析，以比较 Panc1 癌症干样细胞 (Panc1 CSC) 和亲代细胞的分泌蛋白组 线。在 Panc1 CSC 的条件培养基中总共发现 72 种蛋白质上调/下调。通路分析揭示了重要生理通路的调节，包括糖酵解、糖异生和磷酸戊糖。通过 ELISA 免疫测定，我们分析了 PDAC 患者血清中 Panc1 CSC 分泌最高的三种蛋白质（铜蓝蛋白、半乳糖凝集素 3 和 MARCKS）的存在。 ROC 曲线分析表明铜蓝蛋白作为 CA19-9 阴性患者的有希望的标志物。总体而言，我们的研究提供了胰腺 CSC 的系统分泌组分析，揭示了许多参与病理状况（包括癌症分化、侵袭和转移）的分泌蛋白。它们可以作为一个有价值的蛋白质库，从中可以识别生物标志物和治疗靶点。 生物学意义：CSCs的分泌组是癌症进展和分子治疗靶点的生物标志物的丰富库，因此是癌症研究的一个非常有趣的话题。胰腺CSCs的分泌组分析尚未进行。最近，我们课题组证明，利用CSC选择性培养基从亲代细胞系中分离得到的Panc-1 CSCs，为加深对胰腺腺癌生物学的理解具有重要意义。据我们所知，这是第一个胰腺CSC分泌组的蛋白质组学研究。我们进行了基于 iTRAQ 的分析，比较了 Panc1 CSC 和 Panc1 亲本细胞系的分泌组，并鉴定了胰腺癌干细胞以较高水平分泌的总共 43 种蛋白质。我们发现不同重要生理途径（如糖酵解和糖异生、磷酸戊糖途径）的调节以及CSC分泌蛋白（例如72 kDa IV型胶原酶、半乳糖凝集素-3、α-肌动蛋白-4和MARCKS）在癌症分化、侵袭和转移等病理条件中的参与。通过ELISA验证，我们发现MARCKS和 铜蓝蛋白区分对照和 PDAC 患者：此外，ROC 曲线分析表明 MARCKS 不具有诊断准确性，而铜蓝蛋白可能是一种有前途的标记物，仅适用于 CA19-9 阴性的患者。我们认为，我们手稿中报告的发现促进了对胰腺癌肿瘤发生、转移和化疗耐药相关途径的理解，并且还确定了一组 可以从中发现新的候选诊断和治疗生物标志物的蛋白质。 (C) 2016 Elsevier B.V. 保留所有权利。
Emerging research has demonstrated that pancreatic ductal adenocarcinoma (PDAC) contains a sub-population of cancer stem cells (CSCs) characterized by self-renewal, anchorage-independent-growth, long-term proliferation and chemoresistance. The secretome analysis of pancreatic CSCs has not yet been performed, although it may provide insight into tumour/microenvironment interactions and intracellular processes, as well as to identify potential biomarkers.To characterize the secreted proteins of pancreatic CSCs, we performed an iTRAQ-based proteomic analysis to compare the secretomes of Panc1 cancer stem-like cells (Panc1 CSCs) and parental cell line. A total of 72 proteins were found up-/down-regulated in the conditioned medium of Panc1 CSCs. The pathway analysis revealed modulation of vital physiological pathways including glycolysis, gluconeogenesis and pentose phosphate.Through ELISA immunoassays we analysed the presence of the three proteins most highly secreted by Panc1 CSCs (ceruloplasmin, galectin-3, and MARCKS) in sera of PDAC patient. ROC curve analysis suggests ceruloplasmin as promising marker for patients negative for CA19-9.Overall, our study provides a systemic secretome analysis of pancreatic CSCs revealing a number of secreted proteins which participate in pathological conditions including cancer differentiation, invasion and metastasis. They may serve as a valuable pool of proteins from which biomarkers and therapeutic targets can be identified.Biological significance: The secretome of CSCs is a rich reservoir of biomarkers of cancer progression and molecular therapeutic targets, and thus is a topic of great interest for cancer research. The secretome analysis of pancreatic CSCs has not yet been performed.Recently, our group has demonstrated that Panc-1 CSCs isolated from parental cell line by using the CSC selective medium, represent a model of great importance to deepen the understanding of the biology of pancreatic adenocarcinoma.To our knowledge, this is the first proteomic study of pancreatic CSC secretome. We performed an iTRAQ-based analysis to compare the secretomes of Panc1 CSCs and Panc1 parental cell line and identified a total of 43 proteins secreted at higher level by pancreatic cancer stem cells. We found modulation of different vital physiological pathways (such as glycolysis and gluconeogenesis, pentose phosphate pathway) and the involvement of CSC secreted proteins (for example 72 kDa type IV collagenase, galectin-3, alpha-actinin-4, and MARCKS) in pathological conditions including cancer differentiation, invasion and metastasis.By ELISA verification we found that MARCKS and ceruloplasmin discriminate between controls and PDAC patients: in addition ROC curve analyses indicate that MARCKS does not have diagnostic accuracy, while ceruloplasmin could be a promising marker only for patients negative for CA19-9.We think that the findings reported in our manuscript advance the understanding of the pathways implicated in tumourigenesis, metastasis and chemoresistance of pancreatic cancer, and also identify a pool of proteins from which novel candidate diagnostic and therapeutic biomarkers could be discovered. (C) 2016 Elsevier B.V. All rights reserved.