Expression of miR-145 in glial tumors, glioma cell lines and GSCs.

<p>The expression of miR-145 (A) and miR-143 (B) was determined in glial tumors using real-time PCR (p<0.001 for ANOVA F-tests and t-tests for all tumor samples as compared to normal brains). Data from individual human tissues are presented with the median and interquartile range noted. Results are normalized relative to the levels of S12 mRNA and are presented relative to a reference sample. (p<0.005 for all tumor specimens as compared to normal brains). Survival probabilities for patients with GBM expressing low and high levels of miR-145 (C) and miR-143 (D) were determined using Kaplan-Meier estimates and assessed with the log-rank statistic (P = 0.00359, P = 0.00194, respectively). The triangles indicate censored survival time points (i.e. patients alive at last follow-up). The expression of miR-145 in glioma cells as compared to human astrocytes (E) and in GSCs as compared to human neural progenitor cells (F) was determined using real time PCR. Methylation of miR-145 was determined in eight GBM specimens and eight normal brains. An amplicon starting 353 bp upstream pre-miR-145, and ending 78 bp downstream of pre-mir145 promoter, was used for the methylation analysis, covering 12 CpG sites. Zero is the position for the nucleotide starting pre-mir-145. Circles represent CpG site (G). Methylation was determined by Sequenom mass spectrometry detection for each CpG position in the specific amplicon (H). The results are representative of three independent experiments that gave similar results. *P<0.001.