miR-122-5p 通过下调 DUSP4 抑制胃癌细胞迁移和侵袭。
MiR-122-5p inhibits cell migration and invasion in gastric cancer by down-regulating DUSP4.
机构信息
a Department of Laboratory Medicine , Jingjiang People's Hospital , Jingjiang , Jiangsu , China.
b Department of Gastroenterology , Jingjiang People's Hospital , Jingjiang , Jiangsu , China.
出版信息
Cancer Biol Ther. 2018 May 4;19(5):427-435. doi: 10.1080/15384047.2018.1423925. Epub 2018 Mar 6.
OBJECTIVE
To explore the relationship between miR-122-5p and DUSP4 and their effects on gastric cancer (GC) cell mobility and invasiveness.
METHODS
Abnormally expressed miRNAs and mRNAs were analyzed using microarrays. The miR-122-5p and DUSP4 mRNA expression levels in GC tissues and cells were determined by RT-qPCR. The target relationship between miR-122-5p and DUSP4 was validated by dual luciferase reporter assay. GC cell mobility and invasiveness were respectively observed by wound healing assay and transwell invasion assay. Western blot and immunohistochemistry were used for detection of the expressions of DUSP4 protein and MMP2 and MMP9 proteins related to cell invasion and migration. The migration and invasion abilities of gastric cancer cells in vivo were evaluated according to the number of lung metastatic nodules in mice.
RESULTS
The expression of miR-122-5p in GC tissues and cells was significantly down-regulated, whereas DUSP4 expression was up-regulated. Bioinformatics prediction strategies and dual luciferase reporter assay verified the binding sites of miR-122-5p on 3'UTR of DUSP4 and the target relationship between miR-122-5p and DUSP4. Overexpression of miR-122-5p and knockdown of DUSP4 in BGC-823 cells observantly suppressed GC cell mobility and invasiveness, whereas downregulation of miR-122-5p expression promoted cell metastasis. MiR-122-5p inhibited GC cell mobility and invasiveness and pulmonary tumor metastasis via downregulation of DUSP4.
CONCLUSION
MiR-122-5p restrained migration and invasion abilities of GC cells by repressing DUSP4.
目的
探讨 miR-122-5p 与 DUSP4 之间的关系及其对胃癌（GC）细胞迁移和侵袭的影响。
方法
利用微阵列分析异常表达的 miRNAs 和 mRNAs。采用 RT-qPCR 检测 GC 组织和细胞中 miR-122-5p 和 DUSP4mRNA 的表达水平。通过双荧光素酶报告基因实验验证 miR-122-5p 与 DUSP4 之间的靶向关系。通过划痕愈合实验和 Transwell 侵袭实验分别观察 GC 细胞的迁移和侵袭能力。Western blot 和免疫组化检测 DUSP4 蛋白以及与细胞迁移和侵袭相关的 MMP2 和 MMP9 蛋白的表达。根据小鼠肺转移结节的数量评估胃癌细胞体内迁移和侵袭能力。
结果
GC 组织和细胞中 miR-122-5p 的表达明显下调，而 DUSP4 的表达上调。生物信息学预测策略和双荧光素酶报告基因实验验证了 miR-122-5p 与 DUSP4 3'UTR 的结合位点以及 miR-122-5p 与 DUSP4 之间的靶关系。BGC-823 细胞中 miR-122-5p 的过表达和 DUSP4 的敲低显著抑制了 GC 细胞的迁移和侵袭能力，而 miR-122-5p 表达的下调则促进了细胞转移。miR-122-5p 通过下调 DUSP4 抑制 GC 细胞的迁移和侵袭以及肺肿瘤转移。
结论
miR-122-5p 通过抑制 DUSP4 抑制 GC 细胞的迁移和侵袭能力。
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