RASA5 Restoration Suppresses Tumor Cell Invasion and Reverses Cell EMT Phenotype
(A) Matrigel invasion chamber assay of RASA5-expressing MB231 cells. Invaded cells at the lower surface of transwell filter were stained and counted. *, p < 0.05. Scale bar 200 μM. Each bar represented mean ± SEM of three independent experiments with control cells as the baseline using Student's t test.
(B) Morphology changes of HNE1 and MB231 cells stably expressing RASA5. Original magnification, 400x. Scale bar 200 μM.
(C) Ectopic RASA5 expression upregulated E-cadherin, downregulated vimentin, fibronectin and phosphorylation levels of Rac, cofilin, and JNK1/2 by Western blot in HNE1 and MB231 cells. GADPH as a loading control. Graphs represent quantification of Western blots with fold changes compared with controls (right). Data were presented as mean ± SEM of three independent experiments via Student's t test and representative data were shown. *, p < 0.05; **, p < 0.01; ***, p < 0.001.
(D) Expression of EMT markers (E-cadherin, vimentin, and fibronectin) was examined by Western blot in BT549 and T47D cells with RASA5-knockdown through two different siRNAs (siRASA5-1, siRASA5-2). Right panel graphs represent quantification of Western blots with fold changes compared with controls (right). *, p < 0.05; **, p < 0.01. Data were presented as mean ± SEM of three independent experiments via Student's t test and representative data were shown.