© 2021 Marmara University Press.Synthetic cannabinoids have become a significant public health problem in the world because of inexpensive, easy to obtain, difficult to detect by many screening methods, constantly new compounds are produced and marketed, and so on. Despite their effect on ∆9-tetrahydrocannabinol (THC) like properties on cannabinoid receptors, they exert a very strong effect as opposed to THC and could cause serious side effects to death. Studies in the literature on causative epigenetic mechanisms of long-term usage of drugs like cocaine, cannabis, and heroin are available however, few studies have been conducted on synthetic cannabinoids, which are very common in use. Also, recently the endocrine disrupting effects of such substances have come into prominence. We aimed to examine potential effects of AKB48, one of the important synthetic cannabinoids, on cytotoxicity, global methylation and CpG island promoter methylation of cannabinoid receptor genes (CB1 and CB2) and gene expressions of DNA methyltransferase enzymes (DNMT1 and MGMT), CB1 and CB2 genes in prostate adenocarcinoma (PC-3) cells following 24 h exposure. In MTT assay, IC50 value was calculated as 343.8 µM for AKB48 in PC-3 cells. Percentage of 5-methylcytosine (5-mC%) levels were significantly decreased by 27% at 100 μM concentration. No significant alteration was shown in the promoter methylation of CB1 gene compared to the control. These results will contribute to literature for determination of potential effects of synthetic cannabinoids on epigenetic modifications in the tissues of endocrine system.