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. 2021 Mar 24;12:639095. doi: 10.3389/fphar.2021.639095

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Copyright © 2021 Jeong and Kim.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Combination of gefitinib with cilengitide shows enhanced inhibitory effects on TGF-β1-induced EMT expression and Smad signaling. A549 cells were treated with gefitinib (1 μM) and cilengitide (3 μM) individually or in combination, and then incubated with TGF-β1 (5 ng/ml) for 72 h (A, B) or 48 h (C) or 24 h (D, E). The expression of EMT markers (A) and phosphorylation of Smad2/3 (C) were measured by western blot analysis. Actin was used as a loading control. (B) The expression of vimentin was measured by immunocytochemistry. Nuclei were counterstained with DAPI. Scale bars = 100 μm. (D, E) After RNA extraction and cDNA synthesis, we performed qRT-PCR to measure the expression of CDH2 and VIM mRNA using GAPDH as an internal control.^# p < 0.01 vs. control;*p < 0.05, **p < 0.01 vs. the group treated with TGF-β1 only.