Fig. 6

The anti-tumorigenic effect of miR-34a is dependent on MET, and miR-34a-5p inhibits epithelial mesenchymal transition (EMT). a MET vector or control vector (pBABE) were introduced to CAL27 cells and levels of MET mRNA expression were quantified by RT-qPCR. b miR-34a-5p (25 nM) mimic was administrated to the MET overexpressing CAL27 cells and cell proliferation was measured after 24 h using MTT assay. MET signaling was stimulated with HGF (20 ng/ml). c Percentage of early apoptotic cells were determined by Annexin V and PI in MET overexpressing CAL27 cells with and without introduction of miR-34a-5p mimic. MET signaling was stimulated with HGF (20 ng/ml). d Effect of miR-34a-5p on STAT3 protein phosphorylation (shaded line: control; open line: HGF (20 ng/ml); gray line: HGF (20 ng/ml) + miR34a-5p mimic). e, f Relative mRNA expression of epithelial marker (CDH1) and mesenchymal marker (Vimentin) were quantified 48 h after introduction of the miR control mimic or miR-34a-5p mimic by Lipofectamine RNAiMAX in CAL27 cells. RT- qPCR data was normalized using 18 s. g Protein expression of CDH1 and Vimentin were determined after 48 h administration of miR-34a-5p mimic or control mimic in CAL27 cells using western blot. Data represent results of at least three independent experiments. Data is presented as mean ± SD. * indicates p < 0.05. *** < 0.001