Serum and tumor tissue microRNA-23a expression level in glioblastoma patients and the regulation mechanism of ATP5A1 expression by microRNA-23a

Abstract

Objective To test the serum and tumor tissue expression level of microRNA (miRNA, miR)-23a in glioblastoma (GBM) patients and the functional role of miR-23a in GBM. Methods Expression level of serum and tumor tissue miR-23a in GBM patients was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). U87MG, U251, T98G and A172 cells were transfected with miR-23a mimic and the cell proliferation assay was done by CellTiter-Glo® Luminescent Cell Viability Assay kit, at the same time, the tumor growth of U87MG and U251 cells transfected with miR-23a mimic was measured. The expression level of ATP5A1 in GBM tumor tissue was evaluated by RT-qPCR and immunostaining. U87MG, U251, T98G and A172 cells were transfected with miR-23a mimic and the ATP5A1 protein expression was measured by Western blotting. Results The miR-23a expression level in serum (0.40±0.18) and tumor tissue (1.00±0.12) of GBM patients was downregulated compared with health control and tumor adjacent tissue (2.60±0.45 and 1.50±0.56, P=0.005); the miR-23a mimic could suppress the cell growth of U87MG, U251, T98G and A172 cells in vitro (P=0.011), tumor growth of U87MG and U251 cells in vivo, tumor volume of pLKO.1 infected U87MG and U251 was (597.1±95.4) cm3 and (429.5±78.7) cm3, tumor volume of miR-23a mimic infected U87MG and U251 was (153.2±38.6) cm3 and (168.7±33.1) cm3(P=0.008); RT-qPCR and Western blotting results revealed that ATP5A1 (5.4±1.1) was upregulated in GBM tumor tissue, and miR-23a mimic could suppress the protein expression level of ATP5A1 in U87MG, U251, T98G and A172 cells. Conclusion miR-23a could suppress the proliferation and tumor growth of GMB cell lines. Besides it could regulate the protein expression of ATP5A1. Key words: Gioblastoma; MicroRNA-23a; ATP5A1