Zheng Jian, Hernandez Jonathan M, Doussot Alexandre, Bojmar Linda, Zambirinis Constantinos P, Costa-Silva Bruno, van Beek Elke J A H, Mark Milica T, Molina Henrik, Askan Gokce, Basturk Olca, Gonen Mithat, Kingham T Peter, Allen Peter J, D'Angelica Michael I, DeMatteo Ronald P, Lyden David, Jarnagin William R
Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
Children's Cancer and Blood Foundation Laboratories, Department of Pediatrics, Drukier Institute for Children's Health, Meyer Cancer Center, Weill Cornell Medical College, New York, NY, USA; Department of Cell and Developmental Biology, Drukier Institute for Children's Health, Meyer Cancer Center, Weill Cornell Medical College, New York, NY, USA.
HPB (Oxford). 2018 Jul;20(7):597-604. doi: 10.1016/j.hpb.2017.12.010. Epub 2018 Jan 12.
Exosomes are nanovesicles that have been shown to mediate carcinogenesis in pancreatic ductal adenocarcinoma (PDAC). Given the direct communication of pancreatic duct fluid with the tumor and its relative accessibility, we aimed to determine the feasibility of isolating and characterizing exosomes from pancreatic duct fluid.
Pancreatic duct fluid was collected from 26 patients with PDAC (n = 13), intraductal papillary mucinous neoplasm (IPMN) (n = 8) and other benign pancreatic diseases (n = 5) at resection. Exosomes were isolated by serial ultracentrifugation, proteins were identified by mass spectrometry, and their expression was evaluated by immunohistochemistry.
Exosomes were isolated from all specimens with a mean concentration of 5.9 ± 1 × 10 particles/mL and most frequent size of 138 ± 9 nm. Among the top 35 proteins that were significantly associated with PDAC, multiple carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) and extracellular matrix (ECM) proteins were identified. Interestingly, CEACAM 1/5 expression by immunohistochemistry was seen only on tumor epithelia whereas tenascin C positivity was restricted to stroma, suggesting that both tumor and stromal cells contributed to exosomes.
This is the first study showing that exosome isolation is feasible from pancreatic duct fluid, and that exosomal proteins may be utilized to diagnose patients with PDAC.
外泌体是一种纳米囊泡，已被证明在胰腺导管腺癌（PDAC）的致癌过程中起介导作用。鉴于胰管液与肿瘤的直接连通性及其相对易获取性，我们旨在确定从胰管液中分离和鉴定外泌体的可行性。
在手术切除时，从26例PDAC患者（n = 13）、导管内乳头状黏液性肿瘤（IPMN）患者（n = 8）和其他良性胰腺疾病患者（n = 5）中收集胰管液。通过连续超速离心分离外泌体，用质谱法鉴定蛋白质，并通过免疫组织化学评估其表达。
从所有标本中均分离出外泌体，平均浓度为5.9±1×10颗粒/毫升，最常见的大小为138±9纳米。在与PDAC显著相关的前35种蛋白质中，鉴定出多种癌胚抗原相关细胞黏附分子（CEACAMs）和细胞外基质（ECM）蛋白。有趣的是，免疫组织化学检测显示CEACAM 1/5仅在肿瘤上皮细胞上表达，而腱生蛋白C阳性仅限于基质，这表明肿瘤细胞和基质细胞都对外泌体有贡献。
这是第一项表明从胰管液中分离外泌体是可行的研究，并且外泌体蛋白可用于诊断PDAC患者。
