LINC01116 targets miR-520a-3p and affects IL6R to promote the proliferation and migration of osteosarcoma cells through the Jak-stat signaling pathway.

OBJECTIVE: The purpose of this study was to find out the important lncRNA-miRNA-mRNA axis and pathway in osteosarcoma (OS) through bioinformatics analysis and verify the biological functions of lncRNA/miRNA/mRNA in OS through in vitro and in vivo assays.

METHODS: The differential expression mRNAs and lncRNAs were identified through microarray analysis, and the altered pathways were identified by GSEA. The Pearson Coefficient was used to analyze the correlations between mRNAs and lncRNAs. Kaplan-Meier survival analysis was preformed using patient information in GEO database. Their target miRNAs were predicted by Targetscan and miRanda database and confirmed by dual luciferase reporter assay. QRT-PCR were utilized to detected the relative expressions of mRNAs, miRNAs and lncRNAs. The expressions of IL6R protein and pathway related proteins were detected by western blot. OS cell viability, migration and apoptosis were determined through MTT assay, Transwell assay and flow cytometry. Tumor formation in nude mice verified the influence of LINC01116 in vivo.

RESULTS: The Jak-stat signaling pathway was activated in OS tissues. LINC01116 expression was positively correlated with IL6R expression. MiR-520a-3p targeted the 3'-UTR of LINC01116 and IL6R. Lower expression levels of miR-520a-3p significantly correlated with shorter survival of patients. LINC01116 and IL6R were up-regulated while miR-520a-3p was down-regulated in OS. LINC01116 and IL6R promoted the viability and migration of OS cells, while miR-520a-3p acted as a tumor suppressor. MiR-520a-3p inhibitor could rescue the suppressive effects of si-LINC011116 and si-IL6R on OS development. The Jak-stat signaling pathway related proteins were also down-regulated by miR-520a-3p. Down-regulation of LINC01116 inhibited the tumor growth in nude mice.

CONCLUSION: LINC01116 up-regulated IL6R in OS through targeting miR-520a-3p, thus activating the Jak-stat signaling pathway and promoting the progression of OS.