Fig. 2.

Apigenin (API) regulates cellular microfilament bundle and epithelial-to-mesenchymal transition (EMT)-related regulators in non-small cell lung cancer (NSCLC) cells. a Cellular microfilament bundle rearrangements were induced by treating A549 and HCC827 NSCLC cells with API. Cells were treated with 40 μM of vehicle or API for 24 h. Cells were fixed and stained for F-actin by Alexa Fluor 594 Phalloidin (red). Nuclei were counterstained with DAPI (blue). Original magnification, 400×. b A mesenchymal marker, N-cadherin, and the EMT-related transcriptional factors, Snail and Slug, were downregulated in A549 and HCC827 NSCLC cells after 24 h of treatment with API (0~ 40 μM). Quantitative results of indicated protein levels, which were adjusted to the β-actin protein level. c, d Snail family members play a critical role in API-mediated inhibition of the cell migratory ability. Western blot analysis of Snail (c, upper panel) and Slug (c, lower panel) in A549 cells which were transfected with either pLEX-Snail (A549/Snail), pCIneo-Slug (A549/Slug), or their respective controls (A549/Neo) followed by API or vehicle treatment for an additional 24 h. Quantitative results of Snail family protein levels, which were adjusted to the β-actin protein level. Migratory abilities of A549/Snail, A549/Slug, and A549/Neo which were treated with the vehicle or 40 μM API (d). Quantitative results by counting migrated cells in a 100× field. Multiples of differences are presented as the mean ± SD of three independent experiments. Data were analyzed with a one-way ANOVA and Tukey’s post-hoc tests at 95% confidence intervals; different letters represent different levels of significance. Scare bar, 500 μm. e Combined expressions of high Snail and high Slug proteins were correlated with the worst overall survival of patients with lung cancer. The p value refers to a comparison between Snail^high/Slug^high and Snail^low/Slug^low. The lung cancer dataset was retrieved from TCGA