National Key Laboratory of Medical Molecular Biology, Department of Physiology and Pathophysiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences/School of Basic Medicine, Peking Union Medical College, Beijing 100005, China.
Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.
Proteome Sci. 2013 Jul 29;11(1):37. doi: 10.1186/1477-5956-11-37.
Liver perfusates exhibit theoretical advantages regarding the discovery of disease biomarkers because they contain proteins that readily enter the blood-stream, and perfusion preserves the disease state in its natural context. The purpose of the study is to explore the value of liver perfusate proteome in the biomarker discovery of liver diseases.
In this study, 86 differentially expressed proteins were identified in perfusates from isolated rat livers metastasized by Walker-256 tumor cells. Among these proteins, 27 were predicted to be secreted, and 59 were intracellular or membrane proteins. Most of the secretory proteins (70.4%) were decreased in metastasized liver perfusates. The main canonical ingenuity pathway to which these secretory proteins belonged was acute phase response, which indicated that the liver-associated immune reaction was damaged by the metastasis. In contrast, most of the intracellular or membrane proteins (86.4%) exhibited higher relative abundances in the metastasized liver perfusates. Some of these proteins, including Rpl21, Atic, Eif3s2, Echs1, Eps15 and Ywhab, have previously been reported to be involved in cancer genesis and progression. As a member of the 14-3-3 protein family, Ywhab plays a key role in cellular proliferation and oncogenic transformation and has been reported to be involved in the development of breast cancer. Its abundance was elevated by 3.5-fold in the metastasized perfusates. Validation by Western blotting revealed a 3.7-fold increase in the abundance of this protein in metastasized plasma.
These results show that perfusate proteome can be used as an alternative initial resource for biomarker identification, which ultimately requires validation in serum.
肝灌流液在疾病生物标志物的发现方面具有理论优势，因为它们含有易于进入血液的蛋白质，并且灌流在其自然环境中保留了疾病状态。本研究旨在探讨肝灌流蛋白质组在肝脏疾病生物标志物发现中的价值。
在这项研究中，从转移性 Walker-256 肿瘤细胞的大鼠肝脏中分离出的灌流液中鉴定出 86 种差异表达的蛋白质。这些蛋白质中，27 种被预测为分泌蛋白，59 种为细胞内或膜蛋白。大多数分泌蛋白（70.4%）在转移性肝灌流液中减少。这些分泌蛋白主要的经典通路是急性期反应，这表明肝相关免疫反应受到转移的损害。相比之下，大多数细胞内或膜蛋白（86.4%）在转移性肝灌流液中表现出更高的相对丰度。其中一些蛋白质，包括 Rpl21、Atic、Eif3s2、Echs1、Eps15 和 Ywhab，以前曾被报道参与癌症的发生和发展。作为 14-3-3 蛋白家族的成员，Ywhab 在细胞增殖和致癌转化中发挥关键作用，并被报道参与乳腺癌的发展。其在转移性灌流液中的丰度增加了 3.5 倍。Western 印迹验证显示，转移性血浆中该蛋白的丰度增加了 3.7 倍。
这些结果表明，灌流液蛋白质组可以作为生物标志物鉴定的替代初始资源，最终需要在血清中进行验证。
