Figure 2.
VBMECs enhanced NSCLC cell invasion via CX3CL1 signaling-mediated activation of the PI3K/AKT pathway. (A) Western blotting analysis of CX3CL1 protein levels in control (sicon) and CX3CL1-KD (siCX3CL1) VBMECs (Vs), and CX3CR1 protein levels in control (shcon) and CX3CR1-KD (shCX3CR1) A549 cells (A549). (B, C) Quantitative analysis of cell invasion in control (shcon) and CX3CR1-KD (shCX3CR1) A549 cells incubated with or without conditioned culture media of indicated VBMECs (VBMEC CM). Data represent the mean ± SEM (n = 3). **P < 0.01 and ***P < 0.001. (D) Expression levels of EMT markers including Vimentin, E-cadherin, MMP-2, and MMP-9 in control (shcon) and CX3CR1-KD (shCX3CR1) A549 cells co-cultured with or without indicated VBMEC conditioned media measured by western blotting. (E) PI3K and AKT phosphorylation in A549 cells co-cultured with or without VBMEC conditioned media measured by western blotting. (F, G) Quantitative analysis of invasion of A549 cells incubated with or without conditioned culture media of VBMECs treated with the PI3K inhibitor LY294002 or AKT inhibitor MK-2206 2HCI. Data represent the mean ± SEM (n = 3). *P < 0.05 and ***P < 0.001. (H) Expression levels of PI3K, p-PI3K (Ser249), AKT, p-AKT (Ser473), Vimentin, E-cadherin, MMP-2, and MMP-9 in A549 cells co-cultured with indicated VBMECs and treated with LY294002 or MK-2206 2HCI measured by western blotting.