Department of Medical Oncology, Affiliated Wujiang Hospital Of Nantong University, Suzhou, China.
Eur Rev Med Pharmacol Sci. 2018 Nov;22(21):7214-7221. doi: 10.26355/eurrev_201811_16255.
To analyze the role of miR-214-5p in proliferation and metastasis of pancreatic cancer (PC) cells, as well as its underlying mechanism.
30 pairs of PC tissues and adjacent normal tissues were collected in our Department. The expression level of miR-214-5p was detected by quantitative Real-time-polymerase chain reaction (qRT-PCR). Biological information analysis and luciferase report gene assay were used to verify potential target genes of miR-214-5p. Cell counting kit-8 (CCK-8) and transwell methods were applied to observe the interference of miR-214-5p on invasion and migration of PC cells. Western blot (WB) assay was applied to determine the expression changes of Jagged 1 (JAG1) and epithelial-mesenchymal transition (EMT)-related genes in PC cells.
QRT-PCR results showed that the expression level of miR-214-5p is significantly down-regulated in PC tissues and cells. Bioinformatics software and luciferase report gene assay identified that JAG1 is a target gene of miR-214-5p. The negative correlation between protein expressions of miR-214-5p and JAG1 was assessed by Western Blot assay. Furthermore, miR-214-5p could suppress cell proliferation, invasion and migration, and it also blocked the EMT in PC cells in vitro. Meanwhile, JAG1 overexpression reversed the inhibitory effects of miR-214-5p on proliferation, invasion and migration of PC cells.
Overexpressing miR-214-5p could significantly inhibit malignant behavior of PC cells through targeted regulation of JAG1. Thus, miR-214-5p might be a potential therapeutic target for treatment of PC.
分析 miR-214-5p 在胰腺癌细胞增殖和转移中的作用及其潜在机制。
本研究收集了我院 30 对胰腺组织及其相应癌旁正常组织,采用实时荧光定量聚合酶链反应(qRT-PCR)检测 miR-214-5p 的表达水平。采用生物信息学分析和荧光素酶报告基因实验验证 miR-214-5p 的潜在靶基因。通过细胞计数试剂盒-8(CCK-8)和 Transwell 实验观察 miR-214-5p 对胰腺癌细胞侵袭和迁移的干扰作用。Western blot 实验检测胰腺癌细胞中 Jagged 1(JAG1)和上皮间质转化(EMT)相关基因的表达变化。
qRT-PCR 结果显示,miR-214-5p 在胰腺组织和细胞中的表达水平明显下调。生物信息学软件和荧光素酶报告基因实验鉴定出 JAG1 是 miR-214-5p 的靶基因。Western blot 实验评估了 miR-214-5p 与 JAG1 蛋白表达之间的负相关性。此外,miR-214-5p 可抑制胰腺癌细胞的增殖、侵袭和迁移,并在体外阻断 EMT。同时,JAG1 的过表达逆转了 miR-214-5p 对胰腺癌细胞增殖、侵袭和迁移的抑制作用。
过表达 miR-214-5p 通过靶向调控 JAG1 可显著抑制胰腺癌细胞的恶性行为。因此,miR-214-5p 可能成为治疗胰腺的潜在治疗靶点。