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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: MEST promotes lung cancer invasion and metastasis by interacting with VCP to activate NF-κB signaling

Fig. 5

MEST and VCP promote lung cancer migration and invasion via activating the NF-κB pathway. A A549-i8 and H1299-i8 cells were transfected with two anti-VCP siRNAs, and the migration and invasion abilities were determined by using a transwell assay; MMP2 and NF-κB markers (p-IκBα, IκBα, p-p65, and p65) were analyzed by western blot analysis (B). C MEST-overexpressing A549 and H1299 cells were transfected with si-VCP and scramble si-RNA for 24 h. Transwell assay indicates that knockdown of VCP inhibits cell migration and invasion enhanced by MEST in A549 and H1299 cells. Expression of NF-κB markers (p-IκBα, IκBα, p-p65, and p65) were analyzed by western blot analysis (D). E A549-i8 and H1299-i8 were transfected with VCP-HA plasmid or si-MEST for 24 h, as indicated. Transwell assay shows that knockdown of MEST in A549-i8 and H1299-i8 cells decreases cell migration and invasion, and that transfection with VCP-expression plasmid partially restores this effect. The corresponding NF-κB markers (p-IκBα, IκBα, p-p65, and p65) were analyzed by western blot analysis (F). G The indicated cell lines were co-transfected with MMP2 promoter-driven luciferase reporter, with pRL-TK (loading control), as well as with either the indicated plasmids or siRNA. Luciferase activity was measured (n = 3). H Western blot analysis indicates that MMP2 expression is regulated by MEST and VCP. Bars, SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001 compared with control cells unless otherwise indicated. Scale bar, 100 μm

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