2013•Chinese Journal of NeuromedicineRequires access

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

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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

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Available 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

Key concepts: Glioma, Apoptosis, Transfection, Annexin, Molecular biology, MTT assay, Cell growth, Flow cytometry

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