Researches of microRNA-200a on malignant biological behavior of human glioma cells
Xiao-dong Nie, Jinhu Li, Xiaodong Liu
Abstract
Xiao-dong Nie, Jinhu Li, Xiaodong Liu
Abstract
Objective To investigate the effect of microRNA (miRNA, miR)-200a on proliferation, apoptosis, migration and cell cycle of glioma cell line U251 and its possible mechanism. Methods miR-200a mimics (miR-200a group) and nonsense sequences (Mock group) were transfected into human glioma cell line U251 in vitro by liposome transient transfection. Blank group (Blank group) group). The expression of miR-200a was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) after transfection for 48 hours. The cell proliferation ability of the three groups was detected by cell counting kit-8 (CCK-8) method. The cell migration ability was detected by scratches. Cell apoptosis and cell cycle were detected by flow cytometry The expression of ZEB1 and Cyclin D1 protein was detected by Western blotting. Results Compared with the Mock and Blank groups, the relative expression of miR-200a in miR-200a cells was significantly higher than that in the untreated group (P=0.000, 0.000). The cells of miR-200a cells were significantly different (P=0.028, 0.010). The cell migration ability of miR-200a group was significantly decreased (P=0.031, 0.030), the proportion of G0/G1 phase cells in miR-200a group was increased (P=0.011, 0.013), the proportion of S phase cells was significantly decreased (P=0.024, 0.008), and the apoptosis rate of miR-200a group was significantly lower than that of miR-200a group (P=0.001, 0.001). Western blotting showed that the expression of ZEB1 (P=0.042, 0.034) and the expression of Cyclin D1 were significantly down-regulated (P=0.020, 0.013). Conclusion Upregulation of miR-200a can promote glioma cell apoptosis, block its cell cycle progression and inhibit its migration ability, which is expected to be a new target for the treatment of glioma. Key words: Glioma; MicroRNA-200a; Apoptosis; Cell Cycle; Cell Migration
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Objective To investigate the effect of microRNA (miRNA, miR)-200a on proliferation, apoptosis, migration and cell cycle of glioma cell line U251 and its possible mechanism. Methods miR-200a mimics (miR-200a group) and nonsense sequences (Mock group) were transfected into human glioma cell line U251 in vitro by liposome transient transfection. Blank group (Blank group) group). The expression of miR-200a was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) after transfection for 48 hours. The cell proliferation ability of the three groups was detected by cell counting kit-8 (CCK-8) method. The cell migration ability was detected by scratches. Cell apoptosis and cell cycle were detected by flow cytometry The expression of ZEB1 and Cyclin D1 protein was detected by Western blotting. Results Compared with the Mock and Blank groups, the relative expression of miR-200a in miR-200a cells was significantly higher than that in the untreated group (P=0.000, 0.000). The cells of miR-200a cells were significantly different (P=0.028, 0.010). The cell migration ability of miR-200a group was significantly decreased (P=0.031, 0.030), the proportion of G0/G1 phase cells in miR-200a group was increased (P=0.011, 0.013), the proportion of S phase cells was significantly decreased (P=0.024, 0.008), and the apoptosis rate of miR-200a group was significantly lower than that of miR-200a group (P=0.001, 0.001). Western blotting showed that the expression of ZEB1 (P=0.042, 0.034) and the expression of Cyclin D1 were significantly down-regulated (P=0.020, 0.013). Conclusion Upregulation of miR-200a can promote glioma cell apoptosis, block its cell cycle progression and inhibit its migration ability, which is expected to be a new target for the treatment of glioma. Key words: Glioma; MicroRNA-200a; Apoptosis; Cell Cycle; Cell Migration
Key concepts: Cell cycle, Apoptosis, Transfection, Flow cytometry, Blot, Cell growth, Cell counting, Molecular biology