MiR-218 Inhibited Growth And Metabolism Of Human Glioblastoma Cells By Directly Targeting E2F2

Research background: In recently years,the finding of Micro RNA(miRNA)provided a new angle of view on treatment of tumors,including glioma.Mi RNAs are small non-coding RNA that could regulate the expression of related proteins through complementarity to the 3'-untranslated region(3'-UTR)of corresponding m RNA.Although there are a great of studies on miRNA in recently years,only a small number of miRNA has already been investigated.In this study,we focused our attention on miR-218,and its target gene E2F2.E2 F was confirmed,originally,as a cellular factor which could initiate transcription from the viral E2 promoter in adenovirus.Subsequent studies showed that E2 F was very important to the regulation of cell cycle of mammalian cells.From then on,more and more researchers focus on E2 F,the new transcription factor,which maybe play an important role in the progression of cancers.The study showed that miR-218 may be a repressor in glioma though directly targeted to E2F2,and may be a potential therapeutic target for the treatment of glioma.Methods:1.The expression of miRNA and m RNA of glioma cases were downloaded from CGGA data base.2.The expression of miR-218 was detected by q RT-PCR in glioma cases and glioma cell lines.3.Western-blot was employed to test the expression of E2F2 protein in different groups.4.Flow cytometer were used to test the changes of cell cycle in different groups.5.CCK8 assay and Tablet cloning were used to detect the proliferation of cells in different groups.6.The glucose consumption and lactate production of different groups of glioma cell lines were tested by Glucose Colorimetric Assay Kit II and Lactate Colorimetric/Fluorometric Assay Kit according to the manufacturer's instructions.7.Dual luciferase reporter assay were used to identify that E2F2 was a direct target of miR-218.Results1.miR-218 in glioma was decreased compared to the normal brain tissue according to the CGGA database,and low miR-218 expression indicated a short survival time compared to the high miR-218 expression patients.2.Once elevate the expression of miR-218 in glioma cell lines,the glioma cells proliferation were slowed,cell cycle were blocked in G1 phase,and glucose consumption and lactate production were decreased.3.E2F2 in glioma was increased compared to the normal brain tissue according to the CGGA database,and high E2F2 expression indicate a short survival time compared to the low E2F2 expression patients.4.Abrogate the expression of E2F2 in glioma cell lines could suppressed glioma cells growth,blocked cell cycle in G1 phase,and decreased the glucose consumption and lactate production.5.Dual luciferase reporter assay and western-blot showed that E2F2 was a direct target of miR-218.6.Up-regulated the E2F2 expression could partly weaken the effect of miR-218 in glioma cell lines.Conclusions1.The expression of Mi R-218 was decreased in glioma patients,and low miR-218 indicated a short survival time.2.E2F2 in glioma was increased compared to the normal brain tissue,and expression of E2F2 was related to the survival of patients.3.Mi R-218 inhibited growth and metabolism of human glioblastoma cells by directly targeting E2F2,and may be a potential target for the diagnose and treatment of glioma.