Department of Pathology, The Affiliated Southeast Hospital of Xiamen University, Zhangzhou, Fujian 363000, P.R. China.
Department of Gastrointestinal Surgery, Xiamen Humanity Hospital, Xiamen Fujian 361003, P.R. China.
Oncol Rep. 2019 Dec;42(6):2622-2634. doi: 10.3892/or.2019.7387. Epub 2019 Oct 23.
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.
胆管癌（CCA）是一种上皮性癌，其诊断较晚，预后较差。然而，导致 CCA 发生的分子机制尚未完全明确。因此，本研究旨在阐明其中的一些机制。为此，我们采用相对和绝对定量同位素标记（iTRAQ）技术分析了 2 种 CCA 细胞系（TFK1 和 HuCCT1）以及正常胆管上皮细胞系（人肝内胆管上皮细胞，HiBECs）的分泌蛋白。通过差异表达蛋白（DEPs）鉴定和基因本体（GO）功能分类注释及 KEGG 代谢途径图谱分析进行生物学过程分析。使用 RT-qPCR、western blot 分析和免疫组织化学法验证肿瘤蛋白 D52（TPD52）和热休克蛋白家族 DnaJ 成员 B1（DNAJB1）。共鉴定出 778 个 DEPs。经验证后，选择 TPD52 和 DNAJB1 进行进一步分析。在 CCA 细胞系、组织和胆汁样本中，TPD52 和 DNAJB1 的表达水平升高，表明这些蛋白可能参与肿瘤发病机制。此外，TPD52 和 DNAJB1 的表达水平与 CCA 患者的临床参数和预后密切相关。综上所述，本研究结果表明，TPD52 和 DNAJB1 可能作为 CCA 的新型胆汁生物标志物。
