The repressive effect of microRNA-25 inhibitor on human glioma cell growth
张安玲, Kun Wang, 王广秀, 贾志凡, 浦佩玉
Abstract
张安玲, Kun Wang, 王广秀, 贾志凡, 浦佩玉
Abstract
Objective To observe the effect of microRNA (miR-25) inhibitor on glioma cell growth.Methods The miR-25 inhibitor was transfected by liposome. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect miR-25 expression after transfection.The cell cycle kinetics and cell proliferation rate were examined by using flow cytometry and methyl thiazol tetrazolium (MTT) assay,the anchorage-independent cell proliferation was evaluated by soft agar assay,the apoptosis rate was tested by Annexin V staining,and the invasive ability was measured by Transwell.Results The antisense oligonucleotide effectively down-regulated the miR-25 expression in glioma cells.Meanwhile,the proportion of cells in S phase was decreased by about 6%-10%,and the proliferation activity was significantly suppressed (P<0.05 ).Also,the down-regulation of miR-25 promoted glioma cell apoptosis ( P <0.05).However,no significant changes were observed in the number of cells getting through matrigel.Conclusion Suppression of miR-25 expression could inhibit the proliferative ability and induce apoptosis of glioma cells,but had no effect on the invasive ability in glioma cells.This preliminary finding suggests that miR-25 may play an oncogenic role in glioma cell growth,but not invasion. Key words: Glioma; MicroRNA-25; Cell growth; Invasion
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Objective To observe the effect of microRNA (miR-25) inhibitor on glioma cell growth.Methods The miR-25 inhibitor was transfected by liposome. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect miR-25 expression after transfection.The cell cycle kinetics and cell proliferation rate were examined by using flow cytometry and methyl thiazol tetrazolium (MTT) assay,the anchorage-independent cell proliferation was evaluated by soft agar assay,the apoptosis rate was tested by Annexin V staining,and the invasive ability was measured by Transwell.Results The antisense oligonucleotide effectively down-regulated the miR-25 expression in glioma cells.Meanwhile,the proportion of cells in S phase was decreased by about 6%-10%,and the proliferation activity was significantly suppressed (P<0.05 ).Also,the down-regulation of miR-25 promoted glioma cell apoptosis ( P <0.05).However,no significant changes were observed in the number of cells getting through matrigel.Conclusion Suppression of miR-25 expression could inhibit the proliferative ability and induce apoptosis of glioma cells,but had no effect on the invasive ability in glioma cells.This preliminary finding suggests that miR-25 may play an oncogenic role in glioma cell growth,but not invasion. Key words: Glioma; MicroRNA-25; Cell growth; Invasion
Key concepts: Glioma, Apoptosis, Cell growth, Cell cycle, Transfection, Flow cytometry, Annexin, Molecular biology