Department of Oncology, Department of General Surgery Shanghai, China.
Department of Oncology, Department of Pharmacy, Changhai Hospital Affiliated to Second Military Medical University, Shanghai, China.
Carcinogenesis. 2020 Jul 10;41(5):582-590. doi: 10.1093/carcin/bgz186.
Gastric cancer remains one of the most lethal and prevalent malignancies, particularly in China. The majority of patients are diagnosed with gastric cancer at the late stages of the disease. Besides, the high relapse rate also contributes to the high mortality. Therefore, there exists an imperative need for the development of gastric cancer diagnostic techniques as well as novel indicators for gastric cancer metastasis. Exosomes, secreted by a variety of cell types, play critical roles in intercellular communication, which emerge as promising diagnostic biomarkers for gastric cancer. In this study, we present for the first time, at least to the best of our knowledge, the small RNA sequencing spectra of exosomes derived from the gastric cancer patient plasma using next-generation sequencing, focusing on the exploration of metastasis-related biomarkers. The exosomes enriched from patient plasma samples were well characterized by western blotting, transmission electron microscopy and nanoparticle-tracking analysis. In the following bioinformatic analysis of exosomal miRNAs, three candidates were proposed as the biomarkers for metastasis of gastric cancer, namely miR-10b-5p, miR-101-3p and miR-143-5p, for gastric cancer with lymph node metastasis, gastric cancer with ovarian metastasis and gastric cancer with liver metastasis, respectively. RT-qPCR was performed to test the accuracy of these candidates for validation. In conclusion, we successfully isolated and purified exosomes from plasma of patients with gastric cancer and identified several potential exosomal miRNA markers to distinguish gastric cancer patients with various kinds of metastasis.
胃癌仍然是最致命和最常见的恶性肿瘤之一，特别是在中国。大多数患者在疾病晚期被诊断出患有胃癌。此外，高复发率也是导致高死亡率的原因之一。因此，迫切需要开发胃癌诊断技术以及用于胃癌转移的新指标。外泌体由多种细胞类型分泌，在细胞间通讯中发挥着关键作用，是一种很有前途的胃癌诊断生物标志物。在本研究中，我们首次使用下一代测序技术对来自胃癌患者血浆的外泌体进行了小 RNA 测序谱分析，重点探讨了与转移相关的生物标志物。通过 Western blot、透射电子显微镜和纳米颗粒跟踪分析对患者血浆样本中富集的外泌体进行了很好的表征。在接下来的外泌体 miRNA 的生物信息学分析中，提出了三个候选物作为胃癌淋巴结转移、胃癌卵巢转移和胃癌肝转移的转移标志物，分别为 miR-10b-5p、miR-101-3p 和 miR-143-5p。通过 RT-qPCR 验证了这些候选物的准确性。总之，我们成功地从胃癌患者的血浆中分离和纯化了外泌体，并鉴定了几个潜在的外泌体 miRNA 标志物，用于区分具有不同转移类型的胃癌患者。
