Determining The mRNA Which microRNA‐379‐5p Inhibitor Is Targeting in Human Lung A549 Cells

Abstract

MicroRNAs are single‐stranded, non‐coding RNA that are complimentary to messenger RNA and play a role in post‐transcription gene expression and RNA silencing. Understanding the specific mRNA targets of miRNA that are elevated in tumors can provide insight into how cancer develops. The purpose of this experiment was to utilize miR‐379‐5p inhibitor to determine its target genes in human lung cells. Ten target genes for miRNA‐379‐5p were selected from Targetscan and miRDB databases, of which nine were novel and one, Mdm2, was a known target gene used as a positive control. QRT‐PCR primers for target genes were designed. A549 cells were transfected with the miR‐379‐5p inhibitor or a negative control, and then putative target mRNA levels were measured by qRT‐PCR. The Mdm2 positive control gene demonstrated increased levels in the presence of the miR‐379‐5p inhibitor, suggesting the assay was working properly. The putative target gene INSR also showed an increase in mRNA levels in the presence of miR‐379‐5p inhibitor, but a western blot did not show increased expression at the protein level. Other putative target genes examined did not show an increase in mRNA levels in the presence of the miR‐379‐5p inhibitor.