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Fig. 1 | Military Medical Research

Fig. 1

From: Re-expression of DIRAS3 and p53 induces apoptosis and impaired autophagy in head and neck squamous cell carcinoma

Fig. 1

Effects of DIRAS3 and p53 re-expression on the growth and apoptosis of HNSCC cells. a. The expression of DIRAS3 protein in CAL-27 HNSCC cells and noncancerous tongue tissues was determined by western blotting. b. CAL-27 cells were treated with Ad-DIRAS3, rAd-p53, and their combination for 24 h. Ad-GFP was used as a negative control. The morphologic changes were observed using a bright-field microscope. c. Apoptosis after adenoviruses infection was measured by TUNEL assay. The representative images of CAL-27 and SCC-25 cells are shown in the left panels. Green color shown in the images represents positive TUNEL staining, and blue color is from the nuclear dye DAPI. Scale bar, 100 μm. Apoptotic cells in 10 randomly selected fields were counted, and they are shown in the right panels. d. The rate of apoptosis was determined by flow cytometric analysis. Cells with Annexin V+/7-AAD staining were considered to be at an early stage of apoptosis. e. The effects of adenoviruses on the proliferation of CAL-27 and SCC-25 cells were assessed using MTT after 24 h of infection. f. The cell cycle distribution in CAL-27 cells was detected by PI staining and flow cytometric analysis. Representative results from three independent experiments are shown in the upper panels, and the percentage of cells in each phase of the cell cycle is depicted in the lower panels. *P < 0.05; **P < 0.01; ***P < 0.001

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