Linc01124 promotes hepatoblastoma proliferation through the miR-24-3p/PI3K/AKT pathway

Abstract

Children diagnosed with advanced hepatoblastoma (HB) generally have a poor prognosis and are in urgent need of effective treatment strategies. Long non-coding RNAs (lncRNAs) have been found to play crucial roles in various malignant tumors and can be potential targets for tumor diagnosis, treatment and prognosis. However, the role of most lncRNAs in HB pathogenesis is still not well understood. In this study, we investigated the expression of Linc01124 and its effect in HB. Clinical samples of HB patients were collected, and differentially expressed lncRNAs in HB were screened by quantitative real-time polymerase chain reaction (qRT-PCR). The Cell Counting kit-8 and 5-ethynyl-2′-deoxyuridine assay were used to detect cell proliferation in HB cells. The HB cells' apoptosis was assessed by the flow cytometry and terminaldeoxynucleotidyl transferase-mediated nick-end labeling assay. Then micro-RNAs and messenger RNAs targeted by Linc01124 were selected according to miRanda and miRTarbase. The gene pathways regulated by Linc01124 were predicted through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis. The prediction results were subsequently validated using qRT-PCR and Western blotting (WB). A Pearson's correlation analysis was conducted to examine the correlation between hsa-miR-24-3p and Linc01124, IL4, PDGFRB, BRCA1, and NOS3. In addition, HB cells were treated with a Linc01124 inhibitor and PI3K-Akt pathway activator to evaluate the involvement of miR-24-3p/PI3K/AKT in the regulation of HB cell function by Linc01124. Linc01124 was up-regulated in HB cells and tissues. Knockdown of Linc01124 can reduce cell proliferation and increase apoptosis. Bioinformatics analysis predicted that hsa-miR-24-3p could target Linc01124 and 8 genes involved in the PI3K-Akt pathway. qRT-PCR and WB experiments also demonstrated that disrupting Linc01124 expression could increase miR-24-3p expression and inhibit the PI3K/AKT pathway. Meanwhile, the target genes IL4, PDGFRB, BRCA1 and NOS3 expression were also down-regulated by silencing Linc01124. Hsa-miR-24-3p was found to have a negative association with Linc01124, IL4, PDGFRB, BRCA1 and NOS3. PI3K/AKT activator could rescue the decreased proliferation and increased apoptosis induced by Linc01124 knockdown. Linc01124 plays an oncogenic role by improving the activity of the PI3K/AKT pathway, which provides a theoretical basis for future research on the pathogenesis and progression mechanism of HB.