3.6. Cannabis reduced adhesion while it did not modify migration
In cell adhesion experiments, there were no initial differences in the number of dead cells, with 99.9% viability in both the experimental groups when assessed by trypan blue exclusion (data not shown). However, we did observe a decrease in the total number of attached ON cells derived from cannabis users, indicating a weaker adhesive capacity (t= 2.730, p = 0.0293, 3 replicates/cell line, Fig. 5H-J). We also quantified the number of focal adhesions in monolayer cultures of cells immunolabelled with anti-vinculin (Fig. 5K-L) and there were fewer focal adhesions in cells derived from cannabis users (t = 3.859, p = 0.0048, Fig. 5M). Whether the decrease in adhesion was related to an increase in cell migration was determined in a “scratch wound” assay, yet there were no differences in the migration rate of cells from control subjects and cannabis users (percentage closure: t=0.4176, p=0.6872; number of cells: t=0.4619, p=0.6565, Fig. 5N-P). Nevertheless, the proteomic analysis identified differentially expression of several proteins related to cell migration.