Abstract
Long non-coding RNAs (lncRNAs) are emerging as key regulators of neuroblastoma (NB) progression; however, their interplay with MYCN-driven mechanisms remains to be elucidated. The present study aimed to characterize the expression profile of lncRNA RP11-196G11.6 in NB, and to further explore its functional role and mechanism in the pathogenesis of this disease. Transcriptomics data from NB and disseminated tumor cell (DTC) samples (Gene Expression Omnibus; accession no. GSE94035) were analyzed via principal component analysis (PCA), differential expression analysis and CIBERSORT immune profiling. The biological function was assessed using gain- and loss-of-function experiments in IMR-32 (MYCN+) and SH-SY5Y (MYCN-) cells. Dual-luciferase reporter assays were performed to identify the interaction between RP11-196G11.6, microRNA (miR)-376a-3p and RING1 and YY1-binding protein (RYBP). Rescue experiments were performed by co-transfecting RP11-196G11.6-overexpressing cells with a miR-376a-3p mimic to identify the hypothetical regulatory role of RP11-196G11.6 in NB progression in vitro. The present analysis results demonstrated that the PCA could distinguish tumor and DTC samples, with MYCN amplification driving distinct clustering in tumors but not in DTCs. Differential expression analysis based on MYCN+/- in both tumor and DTC groups identified 161 differentially expressed lncRNAs (73 upregulated and 88 downregulated). Notably, RP11-196G11.6 was highly expressed in MYCN+ tumors, and silencing RP11-196G11.6 promoted the viability, migration, invasion and epithelial-mesenchymal transition in MYCN+ cells, whereas RP11-196G11.6 overexpression induced the opposite effects in MYCN- cells. Mechanistically, RP11-196G11.6 directly inhibited miR-376a-3p, which targeted RYBP. Overexpression of miR-376a-3p reversed the tumor suppressor phenotype driven by RP11-196G11.6. In summary, the present study demonstrated that RP11-196G11.6 may inhibit NB progression by sponging miR-376a-3p, leading to the upregulation of RYBP expression and consequently inhibiting NB progression. These findings revealed a novel lncRNA-miRNA axis involved in NB pathogenesis.