Figure 3

miR-146a and miR-181b modulate E-selectin-mediated adhesion of metastatic colon cancer cells to endothelial cells. (A) MiR-146a and miR-181b inhibit E-selectin-dependent adhesion of HT29 colon cancer cells to endothelial cells. Endothelial cells were transfected and stimulated as described in Fig. 1. Calcein AM-stained HT29 metastatic colon cancer cells were added on the tight layer of endothelial cells and incubated for 30 minutes. Non-adhering cells were washed and cells adhering to endothelial cells were determined. To test the E-selectin dependence of the effect of both miRNAs on cancer cells adhesion, anti-E-selectin antibody or MOPC21antibody (1:40) was added to endothelial cells one hour before the addition of HT29 cells. (B) MiR-146a and miR-181b inhibit E-selectin-dependent adhesion of LoVo colon cancer cells to endothelial cells. Experiments were carried out as described in a. Calcein AM-stained LoVo metastatic colon cancer cells were used. The quantification are the mean values of three independent experiments, the error bars represent standard errors of three independent experiments, and the significance was analyzed using a Student’s t-test. The p-values are calculated comparing to the “anti-miR ctrl/IL-1β(/antibody ctrl)” group (*p < 0.05; **p < 0.01). Anti-Esel.: anti-E-selectin antibody.