miR-876-3p 通过靶向 TMED3 调节胃癌细胞对顺铂的耐药性和干细胞样特性。
MiR-876-3p regulates cisplatin resistance and stem cell-like properties of gastric cancer cells by targeting TMED3.
机构信息
Department of Gastrointestinal Surgery, Department of General Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China.
出版信息
J Gastroenterol Hepatol. 2019 Oct;34(10):1711-1719. doi: 10.1111/jgh.14649. Epub 2019 Apr 2.
BACKGROUND AND AIM
Gastric cancer (GC), a prevalent tumor, exerts a major economic burden, and we aimed to explore miR-876-3p's effects on GC and related mechanisms.
METHODS
Cell viability was analyzed via CCK-8 and colony formation assay. Stem cell-like properties were examined via spheroid colony formation assay. mRNA abundance of key genes was analyzed via quantitative polymerase chain reaction. Protein level of TMED3 and stem cell markers was examined by western blot. TargetScan, luciferase, and biotin-miRNA pulldown assay were used to identify miR-876-3p's target.
RESULTS
MiR-876-3p was downregulated in GC, and its mRNA level had negative relationship with cisplatin resistance of GC. Moreover, decreased miR-876-3p expression level suggested poor prognosis of GC patients. MiR-876-3p inhibited drug resistance of cisplatin-resistant cell line SGC-7901/DDP and MKN-45/DDP, as shown by decreased cell viability, IC , and colony formation ability. MiR-876-3p inhibited stem cell-like features and downregulated the expressions of Sox-2, Oct-4, CD133, and CD44 in GC cells. Luciferase and biotin-miRNA pulldown assay confirmed that TMED3 was miR-876-3p's direct target. TMED3 siRNA inhibited miR-876-3p's effects on cisplatin resistance and stem cell-like features of SGC-7901/DDP cells.
CONCLUSION
MiR-876-3p enhanced cisplatin sensitivity and restricted stem cell-like features of GC through targeting TMED3.
背景与目的
胃癌（GC）是一种常见的肿瘤，会造成巨大的经济负担，本研究旨在探讨 miR-876-3p 对 GC 的影响及其相关机制。
方法
通过 CCK-8 和集落形成实验检测细胞活力；通过球体集落形成实验检测肿瘤干细胞样特性；通过定量聚合酶链反应分析关键基因的 mRNA 丰度；通过 Western blot 检测 TMED3 和干细胞标志物的蛋白水平；通过靶基因预测软件、荧光素酶报告基因实验和生物素标记 miRNA 下拉实验鉴定 miR-876-3p 的靶基因。
结果
miR-876-3p 在 GC 中呈低表达，其 mRNA 水平与 GC 对顺铂的耐药性呈负相关。此外，miR-876-3p 表达水平降低提示 GC 患者预后不良。miR-876-3p 抑制了顺铂耐药细胞系 SGC-7901/DDP 和 MKN-45/DDP 的耐药性，表现为细胞活力、IC 50 和集落形成能力降低。miR-876-3p 抑制了 GC 细胞的肿瘤干细胞样特性，并下调了 Sox-2、Oct-4、CD133 和 CD44 的表达。荧光素酶报告基因实验和生物素标记 miRNA 下拉实验证实 TMED3 是 miR-876-3p 的直接靶基因。TMED3 siRNA 抑制了 miR-876-3p 对 SGC-7901/DDP 细胞顺铂耐药性和肿瘤干细胞样特性的影响。
结论
miR-876-3p 通过靶向 TMED3 增强 GC 对顺铂的敏感性并抑制肿瘤干细胞样特征。
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