miR-593 miR-593PLK1miR-593PLK1qRT-PCRWestern blottingPLK1miR-593CCK-8miR-593PLK1 PLK1miRNAmiR-593PLK1miR-593PLK1Western blottingPLK1miR-593 mimicmiR-593 inhibitor 0. control siRNA, PLK1 siRNA, control plasmid or PLK1 plasmid; B: Quantitative analysis of the results of Western blotting; C: Quantitative RT-PCR analysis of PLK1 mRNA expression levels in SW480 cells transfected with control siRNA, PLK1 siRNA, control plasmid or PLK1 plasmid; D: CCK-8 assay of PLK1 protein expression levels in SW480 cells transfected with control siRNA, PLK1 siRNA, control plasmid or PLK1 plasmid. * 0.05, *** 0.001 2.2. PLK1miR-593 3TargetScan[24]miRanda[25]PicTar[26]PLK1miRNAsmiRNsmiR-5933PLK1miRNAmiR-593PLK1 3′-UTR-29.6 kcal/molmiRNAs Rabbit Polyclonal to APOL2 2APLK13’miR-593 2BPLK13’miR-593 mimic 0.01PLK1 3’miR-593miR-593 mimic Open in a separate windows 2 miR-593PLK1 Prediction and dual luciferase reporter plasmid assay to confirm the biding of miR-593 with PLK1 gene. A: Graphical representation of the predicted duplex created by PLK1 3-UTR and miR-593; B: Analysis of luciferase reporter assay. Firefly luciferase reporters made up of either wild-type (WT) or mutant (MUT) miR-593 binding sites in the PLK1 3-UTR were co-transfected into SW480 cells with equivalent doses of the miR-593 mimic and control mimic. ** 0.01 miR-593PLK12miR-593 mimicmiR-593 inhibitor24 hWestern boltmiR-593PLK1miR-593PLK1 0.05 3ABmiR-593miR-593 0.05 3CDPLK1mRNA 0.05 Open in a separate window 3 miR-593PLK1 PLK1 is the direct target gene of miR-593. A, B: Western blot analysis of PLK1 protein levels in SW480 cells and HCT-116 cells after Harpagide transfection with the control mimic, miR-593 mimic, control inhibitor, or miR-593 inhibitor; C: Quantitative RT-PCR analysis of miR-593 level in SW480 cells and HCT-116 cells after transfection; D: Quantitative RT-PCR analysis of PLK1 Harpagide mRNA level in SW480 cells and HCT-116 cells after transfection. The results were expressed as of 3 impartial experiments. * 0.05; ** 0.01, *** 0.001 2.3. miR-593 miR-593 mimicmiR-593 inhibitor24 hCCK-8miR-593miR-593 0.05 4A Open Harpagide in a separate window 4 miR-593PLK1 miR-593 inhibits the proliferation of colon cancer cells by down-regulating PLK1. A: Cell proliferation curve of SW480 cells transfected with the Harpagide control mimic, miR-593 mimic, control inhibitor, or miR-593 inhibitor; B: Cell proliferation curve of SW480 cells transfected with the control mimic plus the control plasmid, the control mimic plus the PLK1 overexpressing plasmid, the miR-593 mimic plus the control plasmid, or the miR-593 mimic plus the PLK1 overexpressing plasmid ( 0.001 control group) 2.4. miR-593PLK1 miR-593PLK1miR-593 mimicPLK124 hCCK-8miR-593PLK1miR-593PLK1 4B 3.? miRNAs[27]miRNAs[28]miR100PLK1[29]miR-593miRNAPLK1[24]PLK1[30]PLK1[31-32]PLK1[33]PLK1miRNAs PLK1miRNAs3PLK1miRNAsmiR-5933[26-28]PLK1miRNACCK-8PLK1[34]miR-593miR-593miR-593PLK1miR-593miRNAmiR- 593 miR-593PLK1PLK1miR-593 mimicPLK13miR-593miR-593PLK1miR-593PLK1miR-593PLK1qRT-PCRPLK1 mRNAmiR-593 mimicmiR-593 inhibitorPLK1mRNAmiR-593PLK1 miR-593 mimicPLK1miR-593 mimicPLK1miR-593PLK1 Western blotqRT-PCRCCK-8miR-593PLK1PLK1miR-593PLK1 Biography ?? E-mail: moc.qq@636063907 Funding Statement 8180265181702275818000822017KJA256 Supported by National Natural Science Foundation of China (81802651, 81702275, 81800082).