Figure 6

MicroRNA-185 reduces the levels of GDM and induces the expression of the promoter-hypermethylated genes. (A) Over-expression of miR-185 reduced the levels of GDM in glioma cell lines. U251, SF126, and SF767 cells were transfected with miR-185 mimics (miR-185M), si-DNMT1, or controls for 48 h and the levels of GDM were determined by HPLC-DAD. Data are expressed as mean ± SD of the relative levels of GDM in each group of cells from three separate experiments. (B) The levels of promoter methylation in the promoter-hypermethylated genes in glioma cell lines. The indicated cells were transfected with miR-185 mimics or control, and the contents of promoter methylation were characterized by MassARRAY system. Data are expressed mean % of the methylated promoters in the indicated genes in those cells. The levels of promoter methylation in these genes were similar to that in unmanipulated cells (data not shown). (C) Quantitative RT-PCR analysis of the relative levels of each gene expression in the indicated cells after transfection with miR-185 mimics, si-DNMT1, or control for 48 h. Data are expressed as mean ± SD of the relative levels of the indicated gene in different cells from three independent experiments. *p< 0.05 vs. control