外泌体 miRNA 表达谱分析及外泌体 miR-4741、miR-32、miR-3149 和 miR-6727 在胃癌进展中的作用。
Exosomal miRNA Expression Profiling and the Roles of Exosomal miR-4741, miR-32, miR-3149, and miR-6727 on Gastric Cancer Progression.
机构信息
Department of Colorectal Surgery, Jinhua Central Hospital of Zhejiang Province, Jinhua, 321000 Zhejiang, China.
出版信息
Biomed Res Int. 2022 Jul 4;2022:1263812. doi: 10.1155/2022/1263812. eCollection 2022.
OBJECTIVE
Accumulated evidence highlights the biological implications of exosomes in gastric cancer. Herein, we conducted the exosomal miRNA expression profiling and identified potential diagnostic markers for gastric cancer.
METHODS
Plasma exosomes were isolated and identified from three gastric cancer patients and three healthy participants. Microarrays of exosomal miRNAs were then analyzed. Differentially expressed exosomal miRNAs were screened with fold - change|≥2.0 and ≤ 0.05. Among them, miR-4741, miR-32, miR-3149, and miR-6727 expressions were verified in tissues and plasma of patients and healthy subjects. ROC curves were conducted for evaluating the diagnostic performance. The roles of miR-32, miR-3149, miR-6727, and miR-4741 on gastric cancer progression were observed by cellular experiments.
RESULTS
Isolated exosomes were well characterized by Western blot and transmission electron microscopy as well as nanoparticle-tracking analyses. According to the microarrays, 142 exosomal miRNAs were upregulated, and 34 were downregulated in gastric cancer than healthy subjects. miR-4741 upregulation and miR-32, miR-3149, and miR-6727 downregulations were found in tissues and plasma of gastric cancer patients. The AUCs of miR-4741, miR-32, miR-3149, and miR-6727 were separately 0.8554, 0.9456, 0.7683, and 0.8923. Upregulated miR-32, miR-3149, and miR-6727 as well as downregulated miR-4741 lowered proliferative, migratory, and invasive capacities as well as elevated apoptotic levels of gastric cancer cells.
CONCLUSION
Our study successfully isolated and verified exosomes from plasma of gastric cancer as well as proposed four exosomal miRNAs that could act as promising diagnostic markers and suppress gastric cancer progression.
目的
越来越多的证据强调了外泌体在胃癌中的生物学意义。在此，我们进行了外泌体 miRNA 表达谱分析，并确定了胃癌的潜在诊断标志物。
方法
从 3 名胃癌患者和 3 名健康参与者的血浆中外泌体进行分离和鉴定。然后分析外泌体 miRNA 的微阵列。用倍数变化|≥2.0 和 ≤ 0.05 筛选差异表达的外泌体 miRNA。其中，miR-4741、miR-32、miR-3149 和 miR-6727 的表达在患者和健康受试者的组织和血浆中得到验证。绘制 ROC 曲线以评估诊断性能。通过细胞实验观察 miR-32、miR-3149、miR-6727 和 miR-4741 对胃癌进展的作用。
结果
Western blot、透射电子显微镜和纳米颗粒跟踪分析均很好地描述了分离的外泌体。根据微阵列，与健康受试者相比，胃癌中外泌体有 142 个上调，34 个下调。在胃癌组织和血浆中发现 miR-4741 上调和 miR-32、miR-3149 和 miR-6727 下调。miR-4741、miR-32、miR-3149 和 miR-6727 的 AUC 分别为 0.8554、0.9456、0.7683 和 0.8923。上调的 miR-32、miR-3149 和 miR-6727 以及下调的 miR-4741 降低了胃癌细胞的增殖、迁移和侵袭能力，并提高了其凋亡水平。
结论
我们的研究成功地从胃癌患者的血浆中分离和验证了外泌体，并提出了四种外泌体 miRNA，它们可以作为有前途的诊断标志物并抑制胃癌的进展。
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