Department of Gynecologic Oncology, Fudan University Shanghai Cancer Center, Shanghai 200032, P.R. China.
Int J Oncol. 2019 May;54(5):1719-1733. doi: 10.3892/ijo.2019.4742. Epub 2019 Mar 7.
Ovarian cancer remains the most lethal type of cancer among all gynecological malignancies. The majority of patients are diagnosed with ovarian cancer at the late stages of the disease. Therefore, there exists an imperative need for the development of early ovarian cancer diagnostic techniques. Exosomes, secreted by various cell types, play pivotal roles in intercellular communication, which emerge as promising diagnostic and prognostic biomarkers for ovarian cancer. In this study, we present for the first time, at least to the best of our knowledge, the proteomics profiling of exosomes derived from the plasma of patients with ovarian cancer via liquid chromatography tandem mass spectrometry (LC‑MS/MS) with tandem mass tagging (TMT). The exosomes enriched from patient plasma samples were characterized by nanoparticle tracking analysis (NTA), dynamic light scattering (DLS), transmission electron microscopy (TEM) and western blot analysis. The size of the plasma exosomes fell into the range of 30 to 100 nm in diameter. The exosomal marker proteins, CD81 and TSG101, were clearly stained in the exosome samples; however, there was no staining for the endoplasmic reticulum protein, calnexin. A total of 294 proteins were identified with all exosome samples. Among these, 225 proteins were detected in both the cancerous and non‑cancerous samples. Apart from universal exosomal proteins, exosomes derived from ovarian cancer patient plasma also contained tumor‑specific proteins relevant to tumorigenesis and metastasis, particularly in epithelial ovarian carcinoma (EOC). Patients with EOC often suffer from coagulation dysfunction. The function of exosomes in coagulation was also examined. Several genes relevant to the coagulation cascade were screened out as promising diagnostic and prognostic factors that may play important roles in ovarian cancer progression and metastasis. On the whole, in this study, we successfully isolated and purified exosomes from plasma of patients with EOC, and identified a potential role of these exosomes in the coagulation cascade, as well as in the diagnosis and prognosis of patients.
卵巢癌仍然是所有妇科恶性肿瘤中最致命的类型。大多数患者在疾病晚期被诊断出患有卵巢癌。因此,迫切需要开发早期卵巢癌诊断技术。外泌体由各种细胞类型分泌,在细胞间通讯中发挥关键作用,它们作为卵巢癌有前途的诊断和预后生物标志物出现。在这项研究中,我们首次通过液相色谱串联质谱法 (LC-MS/MS) 与串联质量标记 (TMT) 展示了来自卵巢癌患者血浆的外泌体的蛋白质组学图谱,至少就我们所知,这是首次展示。通过纳米颗粒跟踪分析 (NTA)、动态光散射 (DLS)、透射电子显微镜 (TEM) 和 Western blot 分析对患者血浆样本中富集的外泌体进行了表征。血浆外泌体的大小落入 30 至 100nm 的直径范围内。外泌体标记蛋白 CD81 和 TSG101 在外泌体样本中清晰染色;然而,内质网蛋白 calnexin 没有染色。总共鉴定出 294 种蛋白质,所有外泌体样本中都检测到 225 种蛋白质。除了通用的外泌体蛋白外,来自卵巢癌患者血浆的外泌体还包含与肿瘤发生和转移相关的肿瘤特异性蛋白,特别是在上皮性卵巢癌 (EOC) 中。EOC 患者常伴有凝血功能障碍。还检查了外泌体在凝血中的功能。筛选出与凝血级联相关的几个基因作为有前途的诊断和预后因素,这些因素可能在卵巢癌进展和转移中发挥重要作用。总的来说,在这项研究中,我们成功地从 EOC 患者的血浆中分离和纯化了外泌体,并确定了这些外泌体在凝血级联中的潜在作用,以及在患者的诊断和预后中的作用。