Department of Interventional Radiology, Zhongshan Hospital, Fudan University, Shanghai, 200032, China; Shanghai Medical Imaging Institute, Shanghai, 200032, China.
J Proteomics. 2013 Oct 8;91:453-65. doi: 10.1016/j.jprot.2013.06.032. Epub 2013 Jul 12.
Histological differentiation is a major pathological parameter associated with poor prognosis in patients with pancreatic adenocarcinoma (PAC) and the molecular signature underlying PAC differentiation may involve key proteins potentially affecting the malignant characters of PAC. We aimed to identify the proteins which could be implicated in PAC prognosis. We used isobaric tags for relative and absolute quantitation (iTRAQ) coupled with two-dimensional liquid chromatography-tandem mass spectrometry to compare protein expression in PAC tissues with different degrees of histological differentiation. A total of 1623 proteins were repeatedly identified by performing the iTRAQ-based experiments twice. Of these, 15 proteins were differentially expressed according to our defined criteria. Myoferlin (MYOF) was selected to validate the proteomic results by western blotting. Immunohistochemistry in a further 154 PAC cases revealed that myoferlin significantly correlated with the degree of histological differentiation (P=0.004), and univariate and multivariate analyses indicated that MYOF is an independent prognostic factor for survival (hazard ratio, 1.540; 95% confidence interval, 1.061-2.234; P=0.023) of patients with PAC after curative surgery. RNA interference-mediated knockdown of MYOF alleviated malignant phenotypes of both primary and metastatic PAC cell lines in vitro and in vivo. Thus, ITRAQ-based quantitative proteomics revealed the prognostic value of MYOF in PAC.
Our results provide the possibility of novel strategies for pancreatic adenocarcinoma management.
组织学分化是与胰腺腺癌 (PAC) 患者预后不良相关的主要病理参数，PAC 分化的分子特征可能涉及潜在影响 PAC 恶性特征的关键蛋白。我们旨在确定可能与 PAC 预后相关的蛋白。我们使用相对和绝对定量同位素标记 (iTRAQ) 与二维液相色谱-串联质谱联用，比较不同组织学分化程度的 PAC 组织中的蛋白表达。通过进行两次基于 iTRAQ 的实验，共鉴定出 1623 种重复蛋白。其中，根据我们定义的标准，有 15 种蛋白差异表达。肌联蛋白 (MYOF) 被选为验证蛋白质组学结果的western blot。在另外 154 例 PAC 病例中进行的免疫组织化学显示，肌联蛋白与组织学分化程度显著相关 (P=0.004)，单因素和多因素分析表明，MYOF 是根治性手术后 PAC 患者生存的独立预后因素 (危险比，1.540；95%置信区间，1.061-2.234；P=0.023)。RNA 干扰介导的 MYOF 敲低减轻了体外和体内原发性和转移性 PAC 细胞系的恶性表型。因此，基于 iTRAQ 的定量蛋白质组学揭示了 MYOF 在 PAC 中的预后价值。
我们的结果为胰腺癌的管理提供了新策略的可能性。
