Regulatory effect of microRNA-148b-3p on long non-coding RNA HOX transcript antisense RNA gene expression in human glioma cell line

Abstract

Objective To investigate the regulation of HOX transcript antisense RNA (HOTAIR) gene by microRNA (miRNA, miR) in human glioma cell line. Methods Bioinformatics analysis was applied to predict the HOTAIR targeted miRNA. MiR-148b-3p mimics, miR-148b-3p inhibitor and their negative controls were bio-synthetized. Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was performed to analyze the regulation of HOTAIR gene expression by miR-148b-3p in human glioma cell line A172 after transferred with miR-148b-3p mimics. Lipofectamine 2000 transfecton and the Dual Luciferase Reporter Gene System were applied to detect the effect of miR-148b-3p on the HOTAIR gene in A172 cell. Results Results of RT-qPCR found that the expression level of HOTAIR in negative control group was 1.06±0.15. 24, 48 and 72 h after miR-148b-3p mimics transfection, the expression level of HOTAIR mRNA was 0.79±0.12, 0.63±0.18 and 0.39±0.06 respectively. 24, 48 and 72 h after miR-148b-3p inhibitor transfection, the expression level of HOTAIR mRNA was 1.24±0.09, 2.70±0.24 and 3.56±0.18 respectively. Compared with the negative control group, miR-148b-3p mimics could decrease the expression of HOTAIR gene (t=2.824, 3.102, 3.648, P 0.05); however, the luciferase activity of A172 cells co-transfected with mimics+ pmirGLO-WT-HOTAIR was significantly decreased compared with A172 cells co-transfected with miR-148b-3p mimics+ pmirGLO-MUT-HOTAIR (t=3.362, P<0.05). Conclusion MiR-148b-3p play a regulatory role of HOTAIR expression by directly targeted combination in human glioma cell lines. HOTAIR is a direct target of miR-148b-3p. Key words: Gliomas; Long non-coding RNA; HOX transcript antisense RNA; MicroRNA-148b-3p