Reduced microRNA 375 in colorectal cancer upregulates metadherin-mediated signaling

Figure 1 Metadherin is a direct target of microRNA 375. A: Sequence alignment of the wild type (WT) and mutated type (MT) microRNA 375 (MIR375) target site in the 3’-UTR of metadherin (MTDH); B: A luciferase reporter plasmid containing WT or MT MTDH 3’-UTR was co-transfected in HCT116 and Caco2 cells with pre-MIR1 as a negative control or pre-MIR375. Results are shown as relative firefly luciferase activity which is standardized to Renilla luciferase activity. Three independent experiments were conducted with duplicates; C: qRT-PCR analysis of MTDH mRNA expression in HCT116 and Caco2 cells. The data are presented as the fold change in MIR375 mimic transfected cells relative to non-transfected cells. Experiment was performed in duplicate and repeated 5 times; D: Cellular MTDH levels in MIR375 mimic-transfected and siMTDH-transfected HCT116, Caco2, and SW48 cells. Three independent experiments were conducted with duplicates. P values were calculated using Student’s t-test (^aP < 0.05, ^bP < 0.01, ^cP < 0.001). MTDH: Metadherin; MIR375: MicroRNA 375; ns: Not significant.