Xu Haisong, Hu Yanxia, Qiu Wusi
Department of Neurosurgery, The Affiliated Hospital of Hangzhou Normal University, Hangzhou 310015, Zhejiang, PR China.
Community Health Service Center of Zha Long Kou Lane, Hangzhou 310004, Zhejiang, PR China.
Biomed Pharmacother. 2017 Mar;87:405-411. doi: 10.1016/j.biopha.2016.12.100. Epub 2017 Jan 6.
The purpose of our study is to clarify the effects of microRNA-129-5p (miR-129-5p) in cellular processes correlated with cancer development and progression of Glioblastoma (GBM) cell by regulating FNDC3B. MiR-129-5p and FNDC3B expression in GBM tissues and tumor adjacent tissues were tested by quantitative real-time PCR. We validated the target relationship between miR-129-5p and FNDC3B by dual luciferase reporter gene system. MTT, colony formation, flow cytometry, Transwell and wound healing assays were used to analyze cell viability, proliferation, apoptosis, invasiveness and migration. The level of FNDC3B protein expression was detected by Western Blot. MiR-129-5p was downregulated in GBM tissues and cell lines, while FNDC3B was upregulated in GBM tissues. The result of luciferase reporter gene assay demonstrated that miR-129-5p could target FNDC3B by binding to the 3' UTR. The overexpression of miR-129-5p or the inhibition of FNDC3B can both inhibit U87 cell viability, proliferation, migration and invasion, while promote cell apoptosis. Our results suggested that miR-129-5p could directly suppress FNDC3B, which might be one of potential mechanisms in inhibiting cell processes including viability, proliferation, migration and invasiveness of U87 cells.
我们研究的目的是通过调节 FNDC3B 来阐明微小 RNA-129-5p(miR-129-5p)在与胶质母细胞瘤(GBM)细胞癌发生和进展相关的细胞过程中的作用。通过定量实时 PCR 检测 GBM 组织和肿瘤邻近组织中 miR-129-5p 和 FNDC3B 的表达。我们通过双荧光素酶报告基因系统验证了 miR-129-5p 与 FNDC3B 之间的靶向关系。采用 MTT、集落形成、流式细胞术、Transwell 和伤口愈合实验分析细胞活力、增殖、凋亡、侵袭和迁移。通过蛋白质免疫印迹法检测 FNDC3B 蛋白表达水平。miR-129-5p 在 GBM 组织和细胞系中表达下调,而 FNDC3B 在 GBM 组织中表达上调。荧光素酶报告基因检测结果表明,miR-129-5p 可通过与 3'UTR 结合靶向 FNDC3B。miR-129-5p 的过表达或 FNDC3B 的抑制均可抑制 U87 细胞的活力、增殖、迁移和侵袭,同时促进细胞凋亡。我们的结果表明,miR-129-5p 可直接抑制 FNDC3B,这可能是抑制 U87 细胞活力、增殖、迁移和侵袭等细胞过程的潜在机制之一。