Suppressive effect of miR-137 on proliferation and apoptosis of U87 glioma cells in vitro
Jun Xia Guo, Zhengqiang Wan, Linshan Fu, Jin Xu, Guan Sun
Abstract
Jun Xia Guo, Zhengqiang Wan, Linshan Fu, Jin Xu, Guan Sun
Abstract
Objective To explore the effect ofmiR-137 on proliferation and apoptosis of U87 glioma cells.Methods The miR-137 mimics were transfected to human glioma cell line U87 as miR-137 transfection group; blank control group and negative control group were also employed in our study.The expression ofmiR-137 was identified by real-time polymerase chain reaction (PCR) after transfection.Methylthiazol tetrazoliu (MTT) assay,flow cytometry-Annexin V/PI assay and fluorescence microscope were employed to detect the cell proliferation and apoptosis.MiR-137 and its relative targeting protein expressions were detected by Westem blotting.Results MTT assay showed that the relative cell survival rates in the blank control group,negative control group and miR-137 transfection group were (105.16±8.57)%,(98.57±8.21)% and (45.84±6.93)%,with significant differences (F=82.157,P=0.000).Annexin V/PI assay showed that the cell apoptosis rates in the blank control group,negative control group and miR-137 transfection group were (4.3±0.63)%,(4.7±0.77)% and (16.6±1.98)%,with significant differences (F=63.837,P=0.000); and apoptotic body was detected by fluorescence microscope.Moreover,CDC42,pERK1/2,AKT and pAKT protein expression levels were inhibited after transfected by miR-137 mimics.Conclusion MiR-137 inhibits proliferation and induces apoptosis of U87 glioma cells,suggesting that miR-137 could be tumor suppressor gene in glioma. Key words: MiR-137; Proliferation; Apoptosis; Glioma
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Objective To explore the effect ofmiR-137 on proliferation and apoptosis of U87 glioma cells.Methods The miR-137 mimics were transfected to human glioma cell line U87 as miR-137 transfection group; blank control group and negative control group were also employed in our study.The expression ofmiR-137 was identified by real-time polymerase chain reaction (PCR) after transfection.Methylthiazol tetrazoliu (MTT) assay,flow cytometry-Annexin V/PI assay and fluorescence microscope were employed to detect the cell proliferation and apoptosis.MiR-137 and its relative targeting protein expressions were detected by Westem blotting.Results MTT assay showed that the relative cell survival rates in the blank control group,negative control group and miR-137 transfection group were (105.16±8.57)%,(98.57±8.21)% and (45.84±6.93)%,with significant differences (F=82.157,P=0.000).Annexin V/PI assay showed that the cell apoptosis rates in the blank control group,negative control group and miR-137 transfection group were (4.3±0.63)%,(4.7±0.77)% and (16.6±1.98)%,with significant differences (F=63.837,P=0.000); and apoptotic body was detected by fluorescence microscope.Moreover,CDC42,pERK1/2,AKT and pAKT protein expression levels were inhibited after transfected by miR-137 mimics.Conclusion MiR-137 inhibits proliferation and induces apoptosis of U87 glioma cells,suggesting that miR-137 could be tumor suppressor gene in glioma. Key words: MiR-137; Proliferation; Apoptosis; Glioma
Key concepts: Glioma, Apoptosis, Transfection, Annexin, Molecular biology, MTT assay, Cell growth, Flow cytometry