微小 RNA-193a-5p 通过调节 NOVA1 在胶质母细胞瘤中发挥肿瘤抑制作用。
MicroRNA-193a-5p exerts a tumor suppressor role in glioblastoma via modulating NOVA1.
机构信息
Department of Neurosurgery, The Affiliated Wenling Hospital of Wenzhou Medical University, Wenling, China.
The Center for Cerebrovascular Disease, The Affiliated Wenling Hospital of Wenzhou Medical University, Wenling, China.
出版信息
J Cell Biochem. 2019 Apr;120(4):6188-6197. doi: 10.1002/jcb.27906. Epub 2018 Oct 10.
BACKGROUND AND OBJECTIVES
Glioblastoma (GBM) is the most common and lethal of intracranial tumors, which is characterized by extensive proliferation and the diffused invasion of tumor cells. MicroRNA-193a-5p (miR-193a-5p) have been demonstrated previously as a functional suppressor in the development and progression of various cancers. The current study aimed to investigate whether miR-193a-5p influences cell proliferation and migration through the mitogen-activated protein kinase/extracellular signal-regulated kinase signaling pathway by targeting neuro-oncological ventral antigen 1 (NOVA1) in glioblastoma.
MATERIALS AND METHODS
The miR-193a-5p expression was detected by quantitative real-time polymerase chain reaction assay in GBM tissues and cell lines. Cell Counting Kit-8 assay, colony formation analysis, wound-healing, and transwell invasion assays were performed to evaluate cell proliferation, colony formation, migration, and invasion, respectively. Western blot analysis and luciferase reporter gene assay were performed to verify the downstream target gene of miR-193a-5p.
RESULTS
The expression of miR-193a-5p was significantly downregulated in GBM tissues and cell lines. Kaplan-Meier analysis showed that patients with low miR-193a-5p expression had a shorter disease-free survival (P < 0.05). Functionally, miR-193a-5p overexpression dramatically suppressed the proliferation, colony formation, migration, and invasion in glioma cells. Bioinformatics prediction and a luciferase assay confirmed that NOVA1 was a direct functional target of miR-193a-5p. Moreover, ectopic expression of NOVA1 could partially reverse the inhibitory effects of miR-193a-5p on glioma cell proliferation, colony formation, migration, and invasion. NOVA1 overexpression abrogated the inhibitory effect of miR-193a-5p on the PTEN/PI3k/AKT pathway.
CONCLUSION
Taken together, our findings suggested that miR-193a-5p functions as a tumor suppressor in glioma cells by directly targeting NOVA1.
背景与目的
胶质母细胞瘤（GBM）是颅内最常见且致命的肿瘤，其特征为肿瘤细胞的广泛增殖和弥漫性浸润。先前的研究表明，微小 RNA-193a-5p（miR-193a-5p）在各种癌症的发生和发展中具有功能抑制作用。本研究旨在通过靶向神经肿瘤性腹侧抗原 1（NOVA1）探讨 miR-193a-5p 是否通过丝裂原活化蛋白激酶/细胞外信号调节激酶信号通路影响胶质母细胞瘤中的细胞增殖和迁移。
材料与方法
采用实时定量聚合酶链反应检测 GBM 组织和细胞系中 miR-193a-5p 的表达。通过细胞计数试剂盒-8 分析、集落形成分析、划痕愈合和 Transwell 侵袭实验分别评估细胞增殖、集落形成、迁移和侵袭。Western blot 分析和荧光素酶报告基因实验验证 miR-193a-5p 的下游靶基因。
结果
miR-193a-5p 在 GBM 组织和细胞系中的表达显著下调。Kaplan-Meier 分析显示，miR-193a-5p 低表达的患者无疾病生存时间较短（P<0.05）。功能上，miR-193a-5p 过表达可显著抑制神经胶质瘤细胞的增殖、集落形成、迁移和侵袭。生物信息学预测和荧光素酶实验证实，NOVA1 是 miR-193a-5p 的直接功能靶基因。此外，NOVA1 的异位表达可部分逆转 miR-193a-5p 对神经胶质瘤细胞增殖、集落形成、迁移和侵袭的抑制作用。NOVA1 过表达可消除 miR-193a-5p 对 PTEN/PI3k/AKT 通路的抑制作用。
结论
综上所述，本研究结果表明，miR-193a-5p 通过直接靶向 NOVA1 在神经胶质瘤细胞中发挥肿瘤抑制作用。
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