Fig. 5.

miR-34a over-expression induces early apoptosis, prevents tumor proliferation, and inhibits tumor growth in vivo. a Representative flow cytometry plot of annexin V (FITC) versus propidium iodide in control mimic treated cells or miR-34a-5p treated HTB-42 cells (25 nM, 48 h). Early apoptotic cells were defined as propidium iodide low and Annexin-V high. b, c The early apoptosis detection assay determined the percentage of early apoptotic cells in 2 different head and neck squamous cell lines (HTB-43 and CAL27 cells), and MET expression was confirmed by western blot. d The level of miR-34a-5p expression was determined by a TaqMan microRNA assay, 12 h post electroporation of miR-34a mimic or control mimic (25 nM). e miR-34a-5p mimic (25 nM or 50 nM) was administered to cells at day 0, and proliferation was measured using MTT reagent to assess the effect of miR-34a on the proliferation of CAL27 cells. f and g Anti-LNA-miR-34a (miR-34a-5p inhibitor) was administered to CAL27 cells and a TaqMan microRNA Assay was used to quantify levels of miR-34a-5p. MET siRNA (20 nM) or scramble siRNA (20 nM) were introduced to CAL27 cells which received miR-34a-5p inhibitor by Lipofectamine RNAiMAX and cell proliferation was quantified by MTT assay. All experiments performed at least in triplicate. Data represent results of at least three independent experiments. Data is presented as mean ± SD.* indicates p < 0.05