Abstract
Objective To examine the expression of microRNA (miRNA, miR)-20a in the glioma cells with temozolomide (TMZ) resistance, and explore the potential molecular mechanisms of TMZ resistance. Methods TMZ resistant glioma cell line, called U251/TMZ cell line, was constructed by the stepwise revulsion with TMZ. The drug resistance and cell viability were detected by cell counting kit-8 (CCK-8) assay. The expression of miR-20a was detected by real-time quantitative polymerase chain reaction (Real-time PCR). The phosphorylated level of mammalian target of rapamycin (mTOR) signal was defined by Western blotting. The level of miR-20a expression in U251/TMZ cells was downregulated by miR-20a inhibitor transfection using lipidosome. Results As cmpared with U251 cells, the expression of miR-20a and the phosphorylated level of mTOR signal were significantly increased in U251/TMZ cells (2.28±0.18 vs. 1.03±0.05, P<0.05). When the expression level of miR-20a in U251/TMZ cells was downregulated by miR-20a inhibitor, the expression of miR-20a and the phosphorylated level of mTOR signal were decreased, and cell viability was decreased in U251/TMZ cells with 50 μmol/L TMZ existing (P<0.01). Conclusion The expression of miR-20a might be contributed to TMZ resistance of glioma cells, which may be related to mTOR signaling way. Key words: Glioma; MicroRNA-20a; Mammalian target of rapamycin; Temozolomide
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