Luciferase reporter assays confirmed a targeting relationship between OIP5-AS1 and miR-410, as well as between miR-410 and Wnt-7b. 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–catenin, GSK-3-pS9, c-Myc and cyclin D1. Furthermore, down-regulation of OIP5-AS1 induced G0/G1 phase cell cycle arrest and apoptosis of glioma AZD3229 Tosylate cells. Inhibitors of miR-410 abolished the biological effects of OIP5-AS1 siRNA in glioma cells. = (= ?0.645, nude mice model, the nude mice in the NC group were not significantly different in terms of tumor volume or weight at various time points compared with those of the Blank group (all We found that silencing OIP5-AS1 using siRNAs in U87 glioma cells inhibited cell growth via effectively suppressing proliferation, invasion and migration capabilities, and promoting apoptosis, as well as inducing G0/G1 phase cell cycle arrest. Consistent with our finding, Naemura et al.  also reported that silencing of OIP5-AS1 modulated the cell cycle and thereby regulated the proliferation of cervical cancer HeLa cells. In the present study, the trend of the miR-410 inhibitors group was observed to be completely opposite to that of the OIP5-AS1 siRNA group, suggesting that blocking miR-410 reversed the inhibitory role of OIP5-AS1 siRNA in terms of growth and metastasis. More importantly, our dual-luciferase assay AZD3229 Tosylate confirmed the targeting relationship between OIP5-AS1 and miR-410, suggesting that OIP5-AS1 may play roles in glioma pathogenesis and progression by modulation of miR-410. Expression of Wnt-7b/-catenin signaling pathway-related proteins was determined to further elucidate the underlying mechanism of OIP5-AS1 in glioma. Expression levels of Wnt-7b, p–catenin, GSK-3-pS9, c-Myc and Cyclin D1 were dramatically down-regulated and the expression of miR-410 was up-regulated in the OIP5-AS1 siRNA group. Nevertheless, cells in the miR-410 inhibitors group showed a completely opposite trend in the changes of these indexes to the OIP5-AS1 siRNA group, showing that inhibition of miR-410 reversed the effect of OIP5-AS1 siRNA to activate the activity of the Wnt-7b/-catenin pathway. Similarly, as illustrated by Shiah et al. , miR-410 attenuated the Wnt–catenin pathway in oral squamous cell carcinoma cells by targeting Wnt-7b, an activator of the Wnt–catenin pathway. Furthermore, Wnt-7b levels were markedly lower in glioma tissues than in nontumor tissues, as illustrated Zhang et al . Notably, the Wnt7b signaling pathway was shown to regulate distinct gliomaCvascular interactions and tumor microenvironments . Using the dual-luciferase reporter assay, we also confirmed that Wnt-7b was indeed the target gene of miR-410, suggesting that silencing OIP5-AS1 may affect growth and metastasis of U87 glioma cells via targeted regulation of Wnt-7b by miR-410. Wnt-7b serves as an important agonist of the Wnt/-catenin signaling pathway [23,24], possibly preventing the phosphorylation and degradation of -catenin induced by GSK-3 inhibition in cytoplasm; the accumulated -catenin would translocate to the nucleus to bind with T-cell factor/lymphoid enhancer factor and then affect the expression of Wnt target genes, including cyclin D1 and c-Myc, eventually promoting tumor pathogenesis AZD3229 Tosylate [25,26]. -Catenin is the core member of the Wnt pathway that has been shown to be highly expressed in high-grade glioma and poorly expressed in low-grade astrocytoma, suggesting that -catenin expression is associated with the degree of malignancy in glioma . There AZD3229 Tosylate was evidence that the knockdown of -catenin greatly changed the growth and cell cycle distribution in glioma, inhibiting the proliferation Rabbit polyclonal to HYAL2 and growth of glioma cells . Moreover, c-Myc is the downstream target gene of the pathway and its enhanced expression was shown to be closely related to the development and progression of glioma . In glioma cells, inhibition of cyclin D1 blocked progression of the cell AZD3229 Tosylate cycle, inhibited proliferation and induced apoptosis [30,31]. Based on all this evidence, we may conclude that OIP5-AS1 siRNA specifically inhibited the Wnt-7b/-catenin pathway and the downstream genes cyclin D1 and c-Myc, resulting in cell cycle arrest, thereby inhibiting cell proliferation, invasion and migration, and promoting apoptosis. Finally, similar results were seen in the nude mice tumorigenesis model also, where tumor quantity and weight had been significantly reduced as well as the development was evidently inhibited in mice treated with OIP5-AS1 siRNA, confirming the inhibitory aftereffect of OIP5-AS1 siRNA on glioma growth even more. Therefore, silencing OIP5-AS1 might inhibit the Wnt-7b/-catenin pathway by up-regulating miR-410, suppressing glioma growth thereby, migration and invasion and marketing apoptosis, deepening knowledge of the molecular system of glioma. Acknowledgments We wish expressing our sincere understanding towards the reviewers because of their constructive comments upon this paper. Abbreviations HEhematoxylinCeosinlncRNAlong.