Figure 3
TFAP2C (transcription factor-activating enhancer-binding protein 2C)-induced TGFBR1 (transforming growth factor-β receptor type 1) upregulation promotes epithelial–mesenchymal transition (EMT) of non-small-cell lung cancer (NSCLC) cells. (a) A three-dimensional (3D) culture model was used to assess the effects of TFAP2C or TGFBR1 knockdown on the morphological changes of NCI-H292 cells. After treatment, the cells were exposed to 4 Gy of radiation, grown in 3D cultures, permeabilized and stained for tubulin (green) and nuclei (blue). (b) The effects of TFAP2C knockdown on the radiation-induced migration of NCI-H292 cells were assessed by a wound-healing assay. Representative images of each group at different time points are shown (top). The graphs show the percentage of the area covered by cells at a specific time point (30 h) from four randomly selected images (bottom). (c) Inhibitory effects of TFAP2C knockdown on the radiation-induced migration of NCI-H292 cells were assessed by a Transwell migration assay. (d) The effects of TFAP2C or TGFBR1 knockdown on the mRNA expression of CDH1, FN1 and VIM in NCI-H292 cells were measured by real-time quantitative RT-PCR (qRT-PCR). (e) Inhibitory effects of TFAP2C knockdown on the protein expression of E-cadherin, fibronectin and vimentin in NCI-H292 and NCI-H838 cells were measured by western blotting. *P<0.05 compared with cells treated with radiation alone; **P<0.05 compared with cells treated with radiation and TFAP2C small interfering RNA (siRNA). Error bars represent±s.e.m. (n=3); one-way analysis of variance (ANOVA), Tukey’s honestly significant difference test.
