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Cancer-Related miRNAs Uncovered by the mirVana miRNA Microarray Platform

ely [12]. We used a Pre-miR miRNA Precursor Molecule and Anti-miR miRNA Inhibitor to one of the miRNAs that were found to be down-regulated in tumor samples to determine if our target site predictions were correct. In these experiments, cultured cells that express undetectable levels of the miRNA were transfected with the Pre-miR Precursor Molecule of the miRNA using siPORT NeoFX Transfection Agent (Ambion). Immunofluorescence assays were used to measure the two putative oncogene targets of the miRNA in the transfected cells. The proteins from both oncogenes were expressed at almost three-fold lower levels in cells transfected with the Pre-miR miRNA than in cells transfected with Pre-miR Negative Control #1 (Ambion) (Figure 5A). In a reciprocal experiment, cells that naturally express high levels of the miRNA were transfected with the Anti-miR miRNA Inhibitor for the miRNA. As expected, the proteins from both oncogenes were higher in cells transfected with the miRNA-specific Anti-miR than in cells transfected with the Anti-miR miRNA Negative Control #1 (Ambion) (Figure 5B). These results are consistent with the model that the miRNA regulates the expression of the two oncogenes.


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Figure 5. Oncogene Levels Modulated in Cells Treated with Cancer-related miRNA Precursor and Inhibitor Molecules.A schematic of each experiment is shown at the left of each panel. (A) Tissue culture cells were transfected with a cancer-related or negative control
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