Expression of miR-145 and miR-449 in U87 Glioblastoma Cells

MicroRNAs are known to play a critical oncogenic role in glioblastoma. Several miRNAs have been shown to play roles in growth and cell cycle control in glioblastoma, and have potential as diagnostic markers or as therapeutic targets. miRNA-145 is overexpressed in glioblastoma and is thought to downregulate srGAP1 (SLIT-ROBO Rho GTPase-activating protein1), which promotes an invasive phenotype. This is in contrast to miRNA-145’s proposed tumor-supressive role in many other cancers (1). miRNA-449 is thought to be a tumor suppressor that interrupts the cell cycle and induces apoptosis via suppression of E2F1, CCND1, and GPR158. Therefore it is highly downregulated in many tumor cells (2,3). Both miRNAs-145 and -449 are the topic of continued inquiry in understanding their expression and their molecular targets in glioma cells. Here we attempt to establish the baseline expression of miRNA-145 and miRNA-449 in the U87 cell line using RT-PCR. Weekly passaging of cells was carried out, followed by RNA isolation and RT-PCR. Baseline average concentrations were established for miRNA-145 and miRNA-449 using a set of serial dilutions and a standard curve. Further experimentation will be required to establish a baseline concentration for these miRNAs in healthy glial cells