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长链非编码 RNA OIP5-AS1 通过调节 miR-410 靶向 Wnt-7b 影响胶质瘤的进展。

Long noncoding RNA OIP5-AS1 targets Wnt-7b to affect glioma progression via modulation of miR-410.

机构信息

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.

出版信息

Biosci Rep. 2019 Jan 3;39(1). doi: 10.1042/BSR20180395. Print 2019 Jan 31.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1bb9/6328889/711b898929d7/bsr-39-bsr20180395-g1.jpg

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