Identification of TPD52 and DNAJB1 as two novel bile biomarkers for cholangiocarcinoma by iTRAQ-based quantitative proteomics analysis
Identification of TPD52 and DNAJB1 as two novel bile biomarkers for cholangiocarcinoma by iTRAQ-based quantitative proteomics analysis
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基于 iTRAQ 的定量蛋白质组学分析将 TPD52 和 DNAJB1 鉴定为胆管癌的两种新型胆汁生物标志物
DOI：
10.3892/or.2019.7387
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发表时间：
2019-12-01
期刊：
影响因子：
4.2
通讯作者：
Chen, Bo
中科院分区：
文献类型：
--
作者：
Ren, Hongyue;Luo, Mingxu;Chen, Bo
胆管癌（CCA）是一种诊断较晚且预后较差的上皮癌。然而，负责CCA的发展的分子机制尚未完全确定。因此，在这项研究中，我们旨在阐明其中的一些机制。为此，进行相对和绝对定量的同量异序标签（iTRAQ）以分析来自2个CCA细胞系TFK 1和HuCCT 1以及来自正常胆管上皮细胞系人肝内胆管上皮细胞（HiBEC）的分泌蛋白。根据基因本体（GO）功能分类注释和KEGG代谢途径图谱分析，鉴定差异表达蛋白（DEPs）并进行生物学过程分析。肿瘤蛋白D52（TPD 52）和DnaJ热休克蛋白家族（Hsp 40）成员B1（DNAJB 1）使用RT-qPCR、蛋白质印迹分析和免疫组织化学进行验证。总共有778个蛋白质被鉴定为DEP。验证后，将TPD 52和DNAJB 1用于进一步分析。TPD 52和DNAJB 1在CCA细胞系、组织和胆汁样品中的表达水平升高，表明这些蛋白可能有助于肿瘤的发病机制。TPD 52和DNAJB 1的表达水平与CCA患者的临床参数和预后密切相关。总体而言，本研究的结果表明，TPD 52和DNAJB 1可能作为新的胆汁生物标志物CCA。
Cholangiocarcinoma (CCA) represents a type of epithelial cancer with a late diagnosis and poor outcome. However, the molecular mechanisms responsible for the development of CCA have not yet been fully identified. Thus, in this study, we aimed to elucidate some of these mechanisms. For this purpose, isobaric tags for relative and absolute quantification (iTRAQ) was performed to analyze the secretory proteins from the 2 CCA cell lines, TFK1 and HuCCT1, as well as from a normal biliary epithelial cell line, human intrahepatic biliary epithelial cells (HiBECs). Differentially expressed proteins (DEPs) were identified and biological process analysis was performed according to the Gene Ontology (GO) functional classification annotation and KEGG metabolic pathway map analysis. tumor protein D52 (TPD52) and DnaJ heat shock protein family (Hsp40) member B1 (DNAJB1) were validated using RT-qPCR, western blot analysis and immunohistochemistry. In total, 778 proteins were identified as DEPs. Following validation, TPD52 and DNAJB1 were used for further analysis. The expression levels of TPD52 and DNAJB1 were elevated in CCA cell lines, tissues and bile samples, suggesting that these proteins may contribute to tumor pathogenesis. In addition, the expression levels of TPD52 and DNAJB1 were found to be closely associated with the clinical parameters and prognosis of patients with CCA. On the whole, the findings of this study indicate that TPD52 and DNAJB1 may serve as novel bile biomarkers for CCA.