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

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

Wei-Li Sun  1 , Tian Kang  2 , Yuan-Yu Wang  3 , Jian-Ping Sun  3 , Chen Li  3 , Hong-Jiang Liu  3 , Yue Yang  3 , Bao-Hua Jiao  4
Affiliations

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

Wei-Li Sun et al. Biosci Rep. .

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. In vivo, 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.

Keywords: Invasion; Long noncoding RNA; Migration; Proliferation; glioma; microRNA.

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Conflict of interest statement

The authors declare that there are no competing interests associated with the manuscript.

Figures

Figure 1
Figure 1. Expression of OIP5-AS1 and miR-410 in glioma tissues and normal tissues measured by qRT-PCR
(A) HE staining used to observe glioma morphology. (B) The relative expression of OIP5-AS1 and miR-410 in glioma tissues and normal tissues. *P<0.05 compared with normal tissues. (C) The correlation analysis between expression levels of OIP5-AS1 and miR-410, P<0.05 indicating a statistically significant difference; (D and E) Relative expression levels of OIP5-AS1 (D) and miR-410 (E) in various grades of glioma tissues; *P<0.05 compared with the low grade glioma tissues.
Figure 2
Figure 2. The targeting relationship among miR-410, OIP5-AS1 and Wnt-7b
(A and B) The online bioinformatics software predicted the binding sites of miR-410 to OIP5-AS1 and Wnt-7b. (C and D) The targeting relationship between miR-410 and OIP5-AS1 and between miR-410 and Wnt-7b verified by the dual-luciferase reporter assay; *P<0.05 compared with the NC group.
Figure 3
Figure 3. Expression of miR-410 and Wnt-7b/β-catenin signaling pathway-related proteins in each group
(A) The expression of Wnt-7b/β-catenin signaling pathway-related proteins assessed by Western blotting. (B) Comparison of the expression of miR-410 in each group. (C–G), Comparison of the expression of Wnt-7b/β-catenin signaling pathway-related proteins in each group; *P<0.05 compared with the Blank group; #P<0.05 compared with the OIP5-AS1 siRNA group; &P<0.05 compared with the miR-410 inhibitors group.
Figure 4
Figure 4. Proliferation of cells in various groups at various time points detected by MTT assay
Figure 5
Figure 5. Comparison of cell cycle and apoptosis rates among groups
Cell cycle (A) and apoptosis (B) among groups detected by flow cytometry. Comparison of cell cycle (C) and apoptosis rates (D) among groups. *P<0.05 compared with the Blank group; #P<0.05 compared with the OIP5-AS1 siRNA group; &P<0.05 compared with the miR-410 inhibitors group.
Figure 6
Figure 6. Comparison of cell invasion and migration among groups
Cell invasion and migration in each group detected by Transwell (A) and wound-healing assays (B). Comparison of the number of invasive cells (C) and wound-healing rate (D) among groups. *P<0.05 compared with the Blank group; #P<0.05 compared with the OIP5-AS1 siRNA group; &P<0.05 compared with the miR-410 inhibitors group.
Figure 7
Figure 7. Effect of OIP5-AS1 and miR-410 on the tumorigenicity of glioma cells in vivo
(A) The tumor volume growth curve of the nude mice in each group (n=8). (B) The subcutaneous tumor weight of the nude mice in each group (n=8); *P<0.05 compared with the Blank group; #P<0.05 compared with the OIP5-AS1 siRNA group. (C) Representative HE staining of the subcutaneous glioma in different groups was shown.
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