Department of Surgery, The University of Hong Kong, Hong Kong, Hong Kong.
Department of Surgery, The University of Hong Kong, Queen Mary Hospital, Hong Kong, Hong Kong.
J Cell Mol Med. 2020 Jun;24(11):6220-6232. doi: 10.1111/jcmm.15253. Epub 2020 May 8.
Exosomes secreted by living cancer cells can regulate metastasis. Exosomal miRNAs can reflect pathological conditions of the original cancer cells. Therefore, we aim to identify exosomal miRNAs as circulating biomarkers for haematogenous metastasis of gastric cancer. Pre-treatment serum samples of eighty-nine patients with stage II/III gastric cancer were collected. Thirty-four of them developed haematogenous metastasis after surgery and the other fifty-five did not. Extraction of exosomes was validated by western blot, transmission electron microscopy and nanoparticle tracking analysis. MiRNA qPCR array was performed in three matched pairs of samples. Internal control was selected from PCR array and validated in the remaining samples. Expressions of exosomal miRNAs were evaluated in the remaining samples by RT-qPCR, as well as in gastric cancer tissue samples and cell culture medium. Expression levels of exosomal miRNAs were analysed with clinical characteristics. The results indicated thirteen up-regulated and six down-regulated miRNAs were found after normalization. MiR-379-5p and miR-410-3p were significantly up-regulated in metastatic patients (P < .01). Higher expression of exosomal miR-379-5p or miR-410-3p showed shorter progression-free survival of the patients (P < .05). It was also found that miR-379-5p and miR-410-3p were down-regulated in gastric cancer tissue samples, while they were significantly up-regulated in gastric cancer cell culture medium compared with cancer cells. In conclusion, exosomal miRNAs are promising circulating biomarkers for prediction of development of haematogenous metastasis after surgery for stage II/III gastric cancer.
活癌细胞分泌的外泌体可以调节转移。外泌体中的 miRNA 可以反映原始癌细胞的病理状况。因此，我们旨在鉴定外泌体 miRNA 作为胃癌血行转移的循环生物标志物。收集了 89 例 II/III 期胃癌患者的术前血清样本。其中 34 例在手术后发生血行转移，其余 55 例未发生。通过 Western blot、透射电镜和纳米颗粒跟踪分析验证了外泌体的提取。在三个配对样本中进行了 miRNA qPCR 阵列分析。从 PCR 阵列中选择内参，并在其余样本中进行验证。通过 RT-qPCR 评估了其余样本中外泌体 miRNA 的表达情况，以及胃癌组织样本和细胞培养物中的表达情况。分析了外泌体 miRNA 的表达水平与临床特征的关系。结果表明，归一化后发现 13 个上调和 6 个下调的 miRNA。转移性患者中 miR-379-5p 和 miR-410-3p 明显上调（P<.01）。外泌体 miR-379-5p 或 miR-410-3p 表达水平较高的患者无进展生存期较短（P<.05）。还发现 miR-379-5p 和 miR-410-3p 在胃癌组织样本中下调，而与癌细胞相比，在胃癌细胞培养物中明显上调。总之，外泌体 miRNA 是预测 II/III 期胃癌手术后血行转移发展的有前途的循环生物标志物。
