Figure 2. Inhibition of proliferation and migration in A549 cells with CB1 and CB2 specific cannabinoids.
A. A549 cells were incubated in culture medium with 100ng/ml EGF (solid bars) and 10% serum (empty bars) for 72 h in the presence of different concentrations of Win55,212-2 and JWH-015 or vehicle control and then analyzed for proliferation by MTT assay. B. A549 and SW-1573 grown in 6-well culture plates were serum starved for 24 h, pretreated with JWH-015 or Win55,212-2 for 30’ prior to stimulation with EGF (10 ng/ml) and incubated for 48 h. Cell scattering was examined by phase-contrast microscopy and photographed. C. Confluent layers of A549 cells were scratched with sterile tips to form wounds and were cultured in the presence of EGF+vehicle or EGF+cannabinoids in presence or absence of GPCR inhibitor Pertussis toxin (PTX). Quantitative analysis of % wound recolonization shows a significant GPCR-mediated inhibition of the EGF-induced chemotaxis by cannabinoids treatment in the A549 cells. Data represent the mean ± SD, representative experiments (n=3) are shown. (*, P<0.05; **, P<0.001; from EGF or serum stimulated, †, P<0.05; ††, P<0.001; from cannabinoid treated)