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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: MicroRNA-486-3p functions as a tumor suppressor in oral cancer by targeting DDR1

Fig. 3

DDR1 is a direct target of miR-486-3p a: Venn diagram of predicted miRNA which targets DDR1 3′-UTR using two independent algorithms (miRNA.org and Targetscan) combined with our patients miRNA array data (GSE45238). b: The luciferase reporter assays with pmirGLO firefly luciferase constructs containing full length of DDR1 3′-UTR in HEK293 cells. The relative luciferase activity of each sample is measured at 48 h after transfection with 20 nM of miRNA mimics (PM-377-3p or PM-486-3p) and normalized to Renilla luciferase activity. The data are represented as mean ± SD; ***p < 0.001 versus control mimics (NC). c: Schematic representation of the putative miR-486-3p binding sequence in the 3′-UTR of DDR1 with wild-type form (DDR1_3′-UTR Wt) and mutant form (DDR1_3’-UTR Mut). The mutated nucleotides are labeled with red color.d: The effect of miRNA mimics (PM-486-3p, 20 nM) on the luciferase activities of the constructs containing the wild-type or mutant-type 3’-UTR in OEC-M1 (left) and TW2.6 (right) cells. The relative luciferase activity of each sampleis measured at 48 h after transfection and normalized to Renilla luciferase activity. The data are represented as mean ± SD; ***p < 0.001 versus control mimics (NC). e: Western blot analysis of DDR1 in OEC-M1 and TW2.6 cells following miR-486-3p mimics (PM-486-3p, 20 nM) transfection for 48 h. f: Validation of miR-486-3p expression by qRT-PCR in 46 of OSCC tumors (T) compared with their own adjacent normal tissues (N). DDR1 expression levels are expressed as the log2 ratios. All data are represented as mean ± SD; ***p < 0.001. g: Correlation analysis of miR-486-3p and DDR1 inhuman OSCC patients (n = 46) by qRT-PCR analysis

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