LncRNA OIP5-AS1通过调节miR-186-5p/KIF14信号轴来促进神经母细胞瘤的增殖

doi:10.3969/j.issn.1006-6233.2024.08.011

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Abstract Objective: To determine the role of long non-coding RNA (LncRNA) Opa-interacting protein 5 antisense RNA 1 (OIP5-AS1) in NB and its potential molecular mechanisms.Methods: Neuroblastoma cell lines SK-N-SH and human neuroblastoma BE2C were cultured in vitro. The expression of OIP5-AS1, miR-186-5p, and kinesin family member 14 (KIF14) was measured by quantitative real-time PCR. Cell proliferation was assessed using the CCK-8 assay, colony formation assay, and EdU incorporation assay. KIF14 protein levels were detected by Western blot. Target genes were predicted using the online database Starbase3.0 and verified by dual-luciferase reporter assay.Results: The expression level of OIP5-AS1 in SK-N-SH cells was significantly higher than that in BE2C cells (1.88±0.14 vs 1.00±0.07, P<0.05). After downregulating OIP5-AS1 expression in the shOIP5-AS1 group, SK-N-SH cell viability, colony formation ability (111.33±15.57 vs 154.67±20.74 vs 149.33±17.01, P<0.05), and DNA synthesis ability (EdU positive cell ratio: 35.09±8.02% vs 67.87±7.21% vs 63.33±7.17%, P<0.05) were weaker than those in the NC group and shCtrl group. Bioinformatics prediction and dual-luciferase reporter assay confirmed the targeting regulation between OIP5-AS1 and miR-186-5p as well as miR-186-5p and KIF14 3'-UTR. Compared with the NC group and shCtrl group, the shOIP5-AS1 group showed downregulated KIF14 mRNA relative expression (0.41±0.09, P<0.05) and KIF14 protein expression (0.20±0.02, P<0.05). Conversely, the shOIP5-AS1+miR-186-5p inhibitor group exhibited increased KIF14 mRNA relative expression and KIF14 protein expression (0.83±0.12 and 0.62±0.04, respectively) compared to the shOIP5-AS1+miR-NC group. RNA-binding protein immunoprecipitation results also showed that, compared with the control (IgG), the Ago2 antibody could enrich OIP5-AS1 and miR-186-5p. Compared with the shOIP5-AS1+miR-NC group, the shOIP5-AS1+miR-186-5p inhibitor group had significantly increased cell viability and colony formation ability (180.0±11.36 vs 136.0±14.53, P<0.05). In rescue experiments, compared with the shOIP5-AS1+miR-186-5p inhibitor+siNC group, the shOIP5-AS1+miR-186-5p inhibitor+siKIF14 group showed significantly decreased cell viability and colony formation ability (139.33±8.96 vs183.03±18.0, P<0.05).Conclusion: The expression level of OIP5-AS1 is significantly upregulated in SK-N-SH cells and acts as a competing endogenous RNA (ceRNA) to competitively inhibit miR-186-5p, thereby upregulating KIF14 expression and promoting NB progression.

Key words： Neuroblastoma LncRNA Opa-interacting protein 5 antisense RNA 1 Kinesin family member 14

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	AN Yanxiao
	JIAO Hanliang
	QI Yanwei
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AN Yanxiao,JIAO Hanliang,QI Yanwei, et al. Effect of LncRNA OIP5-AS1 on Proliferation of Neuroblastoma by Regulating the miR-186-5p/KIF14 Signaling Axis[J]. HeBei Med, 2024, 30(8): 1290-1296.

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http://www.hbyxzzs.cn/EN/10.3969/j.issn.1006-6233.2024.08.011 OR http://www.hbyxzzs.cn/EN/Y2024/V30/I8/1290

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