Autophagy is a catabolic process involved in homeostatic and regulated cellular protein recycling and degradation via the lysosomal degradation pathway. Emerging data associates Crohn's Disease (CD) with an impaired ATG16L1 autophagy gene. Increased activity in the endocannabinoid system and up-regulation of suppressor of cytokine signalling (SOCS)-3 protein expression are evident in inflamed intestine. We accessed the impact of phyto-cannabinoid (CBD), synthetic cannabinoid (ACEA) and endocannabinoid (AEA) on autophagosome formation, and investigated the mechanisms involved. Our findings show that all three cannabinoids induce autophagy in a dose-dependent manner in fully differentiated CaCo2 cells. ACEA and AEA induced canonical autophagy, which was cannabinoid receptor (CNR)-l mediated. In contrast, CBD-induced autophagy is partially non-canonical and not CNR1 receptor mediated. Functionally, all three cannabinoids reduce SOCS3 protein expression. Blocking of autophagy reversed the cannabinoid-induced effect. In conclusion, CBD may have potential therapeutic application in CD where functional CNR1 receptor or autophagy is compromised and the regulatory protein, SOCS3, is itself regulated by the autophagy pathway.