Effect of LncRNA OIP5-AS1 on Proliferation of Neuroblastoma by Regulating the miR-186-5p/KIF14 Signaling Axis
Objective:To determine the role of long non-coding RNA(LncRNA)Opa-interacting pro-tein 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 a-bility(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 tar-geting regulation between OIP5-AS1 and miR-186-5p as well as miR-186-5p and KIF14 3'-UTR.Com-pared with the NC group and shCtrl group,the shOIP5-AS1 group showed downregulated KIF14 mRNA rela-tive 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 colo-ny 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.
NeuroblastomaLncRNAOpa-interacting protein 5 antisense RNA 1Kinesin family member 14
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