title: Colon carcinogenesis is inhibited by the TRPM8 antagonist cannabigerol, a Cannabis-derived non-psychotropic cannabinoid.
abstract: Cannabigerol (CBG) is a safe non-psychotropic Cannabis-derived cannabinoid (CB), which interacts with specific targets involved in carcinogenesis. Specifically, CBG potently blocks transient receptor potential (TRP) M8 (TRPM8), activates TRPA1, TRPV1 and TRPV2 channels, blocks 5-hydroxytryptamine receptor 1A (5-HT1A) receptors and inhibits the reuptake of endocannabinoids. Here, we investigated whether CBG protects against colon tumourigenesis. Cell growth was evaluated in colorectal cancer (CRC) cells using the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide and 3-amino-7-dimethylamino-2-methylphenazine hydrochloride assays; apoptosis was examined by histology and by assessing caspase 3/7 activity; reactive oxygen species (ROS) production by a fluorescent probe; CB receptors, TRP and CCAAT/enhancer-binding protein homologous protein (CHOP) messenger RNA (mRNA) expression were quantified by reverse transcription-polymerase chain reaction; small hairpin RNA-vector silencing of TRPM8 was performed by electroporation. The in vivo antineoplastic effect of CBG was assessed using mouse models of colon cancer. CRC cells expressed TRPM8, CB1, CB2, 5-HT1A receptors, TRPA1, TRPV1 and TRPV2 mRNA. CBG promoted apoptosis, stimulated ROS production, upregulated CHOP mRNA and reduced cell growth in CRC cells. CBG effect on cell growth was independent from TRPA1, TRPV1 and TRPV2 channels activation, was further increased by a CB2 receptor antagonist, and mimicked by other TRPM8 channel blockers but not by a 5-HT1A antagonist. Furthermore, the effect of CBG on cell growth and on CHOP mRNA expression was reduced in TRPM8 silenced cells. In vivo, CBG inhibited the growth of xenograft tumours as well as chemically induced colon carcinogenesis. CBG hampers colon cancer progression in vivo and selectively inhibits the growth of CRC cells, an effect shared by other TRPM8 antagonists. CBG should be considered translationally in CRC prevention and cure.
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via D. Montesano 49, 80131 Naples, Italy, Department of Diagnostic Services (Anatomy and Pathologic Histology Service), Ospedale dei Pellegrini, ASL 1, 80135 Naples, Italy, Institute of Biomolecular Chemistry, National Research Council, Via Campi Flegrei 34, 80078 Pozzuoli, Naples, Italy and Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, 80131 Naples, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Borrelli
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via D. Montesano 49, 80131 Naples, Italy, Department of Diagnostic Services (Anatomy and Pathologic Histology Service), Ospedale dei Pellegrini, ASL 1, 80135 Naples, Italy, Institute of Biomolecular Chemistry, National Research Council, Via Campi Flegrei 34, 80078 Pozzuoli, Naples, Italy and Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, 80131 Naples, Italy.
authors.forename: Ester
authors.initials: E
authors.lastname: Pagano
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via D. Montesano 49, 80131 Naples, Italy, Department of Diagnostic Services (Anatomy and Pathologic Histology Service), Ospedale dei Pellegrini, ASL 1, 80135 Naples, Italy, Institute of Biomolecular Chemistry, National Research Council, Via Campi Flegrei 34, 80078 Pozzuoli, Naples, Italy and Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, 80131 Naples, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Romano
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via D. Montesano 49, 80131 Naples, Italy, Department of Diagnostic Services (Anatomy and Pathologic Histology Service), Ospedale dei Pellegrini, ASL 1, 80135 Naples, Italy, Institute of Biomolecular Chemistry, National Research Council, Via Campi Flegrei 34, 80078 Pozzuoli, Naples, Italy and Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, 80131 Naples, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Panzera
authors.affiliation: Department of Diagnostic Services (Anatomy and Pathologic Histology Service), Ospedale dei Pellegrini, ASL 1, 80135 Naples, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Maiello
authors.affiliation: Department of Diagnostic Services (Anatomy and Pathologic Histology Service), Ospedale dei Pellegrini, ASL 1, 80135 Naples, Italy.
authors.forename: Diana
authors.initials: D
authors.lastname: Coppola
authors.affiliation: Institute of Biomolecular Chemistry, National Research Council, Via Campi Flegrei 34, 80078 Pozzuoli, Naples, Italy and.
authors.forename: Luciano
authors.initials: L
authors.lastname: De Petrocellis
authors.affiliation: Institute of Biomolecular Chemistry, National Research Council, Via Campi Flegrei 34, 80078 Pozzuoli, Naples, Italy and.
authors.forename: Lorena
authors.initials: L
authors.lastname: Buono
authors.affiliation: Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, 80131 Naples, Italy.
authors.forename: Pierangelo
authors.initials: P
authors.lastname: Orlando
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via D. Montesano 49, 80131 Naples, Italy, Department of Diagnostic Services (Anatomy and Pathologic Histology Service), Ospedale dei Pellegrini, ASL 1, 80135 Naples, Italy, Institute of Biomolecular Chemistry, National Research Council, Via Campi Flegrei 34, 80078 Pozzuoli, Naples, Italy and Institute of Protein Biochemistry, National Research Council, Via P. Castellino 111, 80131 Naples, Italy aaizzo@unina.it.
authors.forename: Angelo A
authors.initials: AA
authors.lastname: Izzo
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001397:Azoxymethane / Q000633:toxicity; D015153:Blotting, Western; D002186:Cannabinoids / Q000494:pharmacology*; D002188:Cannabis / Q000737:chemistry; D002273:Carcinogens / Q000633:toxicity; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D003110:Colonic Neoplasms / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology; D005434:Flow Cytometry; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008819:Mice, Nude; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050053:TRPM Cation Channels / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D002186:Cannabinoids; D002273:Carcinogens; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D050053:TRPM Cation Channels; C492297:TRPM8 protein, mouse; C037036:cannabigerol; D001397:Azoxymethane