Potential Role of Long Non-Coding RNA ANRIL in Pediatric Medulloblastoma Through Promotion on Proliferation and Migration by Targeting miR-323.

Medulloblastoma (MB) is a common primary tumor of the central nervous system diagnosed in pediatric patients. Survivors of MB are frequently accompanied by severe side effects, thus it is urgent to explore novel therapeutic target. First of all, the level of antisense non-coding RNA in the INK4 locus (ANRIL) in normal cerebellum and DAOY cells were evaluated. Then, the effect of ANRIL on DAOY cell proliferation, migration, and apoptosis were tested. Then, qRT-PCR was performed to explore whether ANRIL acted as a sponge of miR-323. The effects of miR-323 inhibition on ANRIL silence-induced alterations of cell proliferation, migration, and apoptosis were estimated. Predicted by TargetScan, the possible target gene of miR-323 was screened, followed by validation with luciferase assay. The abnormally expressed BRI3 on cell proliferation, migration, apoptosis, and signaling pathways were all evaluated. ANRIL was up-regulated in MB and its silence significantly lowered cell viability and migration but promoted cell apoptosis. ANRIL acted as a sponge of miR-323 and its silence functioned through up-regulating miR-323. BRI3 and CDK6 were target genes of miR-323 and the effect of BRI3 on DAOY cells was the same as ANRIL. Moreover, ANRIL suppression could reduce phosphorylated levels of p38 MAPK, ERK and AKT, and inhibit Wnt signaling pathway through positively regulating BRI3. ANRIL inhibition repressed cell proliferation and migration but promoted cell apoptosis through miR-323-mediated regulation of BRI3, which could activate p38 MAPK, ERK, and AKT as well as Wnt signaling pathway. J. Cell. Biochem. 118: 4735-4744, 2017.