microrna-195对人脑胶质瘤干细胞的作用word论文.docxVIP

Effects of MicroRNA-195 on Human Glioma Stem CellsAbstractObjective: To detect the expression of microRNA-195 (miR-195) in human glioma stem cells and explore the mechanism of regulation of miR-195 in human glioma stem cells.Methods: First we cultured the glioblastoma (GBM) cell line U87 with Neuro stem cells (NSCs) medium and obtained the tumor spheres of Glioma stem cells(GSCs) named as U87-S, whose properties were detected by sphere formation assays. Then the expression of miR-195 in U87 cells and U87-S cells were detected by qRT-PCR. Thereafter miR-195 mimics and scrambled control oligonucleotides were transfected into GSCs by using Lipofectamine RNAi MAX. The transfection condition of miR-195 group was detected with a fluorescence microscope, and qRT-PCR was applied to measure the expression level of miR-195 in transfected group, the negative control group and the U87-S cells. Then the GSCs’ self-renewal ability was tested by sphere formation assay, and immunofluorescence analysis was carried out to examine the differentiation condition of the cells of the two groups. Cell proliferation was determined by Cell Counting Kit-8(CCK-8). The apoptosis ratio was analyzed by FACS. Data was expressed as mean ± the standard error. Statistical analysis was performed with the SPSS13.0 software.One-way ANOVA was used for the comparison of several groups’ mean, and LSD－t test wasused to analyze the variance among the groups in the number of pairwise comparisons．P 0.05 was considered significant.Results: 1. GSCs was achieved by culturing U87 cells in NSCs medium, named U87-S. 2. Single cell dissociated from GSCs spheres had the ability to form secondary spheres. 3. The result of qRT-PCR showed that expression of miR-195 was significantly decreased in U87-S cells compared with U87 cells (p0.05), and transfected group cells had a higher level of miR-195 than negative control group (p0.05). 4. The transfected efficiency of miR-195 mimicsin the transfected group was above 9