Department of Rehabilitation, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei 050000, P.R. China.
Department of Pediatrics, the First Hospital of Shijiazhuang, Shijiazhuang 050011, China.
Biosci Rep. 2019 Jan 3;39(1). doi: 10.1042/BSR20180395. Print 2019 Jan 31.
The present study was undertaken to investigate the underlying mechanisms of long noncoding RNA OIP5-AS1 via regulating miR-410 to modulate Wnt-7b in the progression of glioma. To address this problem, we measured the expression of OIP5-AS1 and miR-410 in glioma tissues by qRT-PCR. Glioma U87 cells were transfected with OIP5-AS1 siRNA or miR-410 inhibitors. The targeting relationships among miR-410, OIP5-AS1 and Wnt-7b were verified by luciferase reporter assays. Western blotting was employed to determine the expression of Wnt-7b/β-catenin pathway-related proteins, while MTT, flow cytometry, Transwell assays and wound-healing assays were used to measure the biological characteristics of glioma cells. The results showed that OIP5-AS1 expression was higher and miR-410 was lower in glioma tissues. Luciferase reporter assays confirmed a targeting relationship between OIP5-AS1 and miR-410, as well as between miR-410 and Wnt-7b. Silencing OIP5-AS1 reduced cell proliferation, invasion and migration of glioma U87 cells and led to depressed expression levels of miR-410, Wnt-7b, p-β-catenin, GSK-3β-pS9, c-Myc and cyclin D1. Furthermore, down-regulation of OIP5-AS1 induced G0/G1 phase cell cycle arrest and apoptosis of glioma cells. Inhibitors of miR-410 abolished the biological effects of OIP5-AS1 siRNA in glioma cells. , OIP5-AS1 knockdown also inhibited tumor growth. Taken together, this research suggested that silencing OIP5-AS1 may specifically block the Wnt-7b/β-catenin pathway via targeted up-regulating miR-410, thereby inhibiting growth, invasion and migration while promoting apoptosis in glioma cells.
本研究旨在通过调控长非编码 RNA OIP5-AS1 介导的 miR-410 来研究其在胶质瘤进展中的潜在机制，从而调节 Wnt-7b。为了解决这个问题，我们通过 qRT-PCR 测量了胶质瘤组织中 OIP5-AS1 和 miR-410 的表达。用 OIP5-AS1 siRNA 或 miR-410 抑制剂转染胶质瘤 U87 细胞。通过荧光素酶报告基因实验验证了 miR-410、OIP5-AS1 与 Wnt-7b 之间的靶向关系。采用 Western blot 检测 Wnt-7b/β-catenin 通路相关蛋白的表达，通过 MTT、流式细胞术、Transwell 实验和划痕愈合实验检测胶质瘤细胞的生物学特性。结果表明，胶质瘤组织中 OIP5-AS1 表达升高，miR-410 表达降低。荧光素酶报告基因实验证实 OIP5-AS1 与 miR-410 之间以及 miR-410 与 Wnt-7b 之间存在靶向关系。沉默 OIP5-AS1 可降低胶质瘤 U87 细胞的增殖、侵袭和迁移能力，并导致 miR-410、Wnt-7b、p-β-catenin、GSK-3β-pS9、c-Myc 和 cyclin D1 的表达水平降低。此外，下调 OIP5-AS1 可诱导胶质瘤细胞发生 G0/G1 期细胞周期阻滞和凋亡。miR-410 抑制剂可消除 OIP5-AS1 siRNA 在胶质瘤细胞中的生物学效应。此外，OIP5-AS1 敲低还抑制了肿瘤的生长。综上所述，本研究表明沉默 OIP5-AS1 可能通过靶向上调 miR-410 特异性阻断 Wnt-7b/β-catenin 通路，从而抑制胶质瘤细胞的生长、侵袭和迁移，同时促进其凋亡。
