Fatty acid amide hydrolase (FAAH) inhibitor PF-3845 reduces viability, migration and invasiveness of human colon adenocarcinoma Colo-205 cell line: an in vitro study

摘要

Earlier reports suggest that the endocannabinoids mayplay a role of endogenous tumor growth modulators.In this study, we investigated whether inhibition of theenzymes involved in the synthesis and degradation ofendocannabinoids may reduce colorectal cancer cellinvasion and migration. The human colon adenocarcinomaColo-205 cells were incubated with PF-3845, JZL-184 and RHC-80267 (fatty acid amide hydrolase (FAAH),mono- (MAGL) and diacylglycerol lipase (DAGL) inhibitors,respectively) for 48 h. The MTT colorimetric assaywas performed to quantify cell viability. Next, Colo-205cells were incubated with PF-3845 alone or with PF-3845together with selected antagonists: AM 251, AM 630, SB366791, RN 1734 and G-15 (CB1, CB2, TRPV1, TRPV4 andGPR30 antagonists, respectively). Western blot assay wasapplied to identify the changes in CB1 and CB2 receptorexpression. Migration and invasion assays were employedto characterize the effect of PF-3845 on colorectalcancer cell invasion. We found that of all the inhibitorsused, the FAAH inhibitor PF-3845 reduced the Colo-205 cell line viability the most effectively (IC50=52.55μM). We also showed that the effect of decreased cellviability was enhanced when Colo-205 cells were incubatedwith PF-3845 and RN-1734, a TRPV4 antagonist(IC50=30.54 μM). Western blot assay revealed significantlydecreased CB1 receptor expression levels, while CB2 expressionwas increased in response to PF-3845 whencompared to control. Furthermore, PF-3845 inhibited migrationand invasion of Colo-205 cell line. These resultssuggest that pharmacological inhibition of FAAH andconsequent enhancement of the endocannabinoid levelsmay reduce the colorectal cancer growth and progression.