AM251 induces apoptosis and G2/M cell cycle arrest in A375 human melanoma cells
Year: 2015
Authors: Carpi, S., Fogli, S., Romanini, A., Pellegrino, M., Adinolfi, B., Podesta, A., Costa B., Da Pozzo E., Martini, C., Breschi M.C., Nieri, P.
Autors Affiliation: University of Pisa, Deptments of Pharmacy, I-56126 Pisa, Italy; University of Pisa, Dept. Translational Research and New Technologies in Medicine and Surgery, I-56126 Pisa, Italy; University of Pisa, Dept. Veterinary Sciences, I-56126 Pisa, Italy; University Hospital of Pisa, Medical Oncology Unit, Pisa, Italy
Abstract: Human cutaneous melanoma is an aggressive and chemotherapy-resistant type of cancer. AM251 is a cannabinoid type 1 (CB1) receptor antagonist/inverse agonist with off-target antitumor activity against pancreatic and colon cancer cells. The current study aimed to characterize the in-vitro antimelanoma activity of AM251. The BRAF V600E mutant melanoma cell line, A375, was used as an in-vitro model system. Characterization tools included a cell viability assay, nuclear morphology assessment, gene expression, western blot, flow cytometry with Annexin V-FITC/7-AAD double staining, cell cycle analyses, and measurements of changes in intracellular cAMP and calcium concentrations. AM251 exerted a marked cytotoxic effect against A375 human melanoma cells with potency comparable with that observed for cisplatin without significant changes in the human dermal fibroblasts viability. AM251, at a concentration that approximates the IC50, downregulated genes encoding antiapoptotic proteins (BCL2 and survivin) and increased transcription levels of proapoptotic BAX, induced alteration of Annexin V reactivity, DNA fragmentation, chromatin condensation in the cell nuclei, and G2/M phase arrest.AM251 also induced a 40% increase in the basal cAMP levels, but it did not affect intracellular calcium concentrations. The involvement of GPR55, TRPA1, and COX-2 in the AM251 mechanism of action was excluded. The combination of AM251 with celecoxib produced a synergistic antitumor activity, although the mechanism underlying this effect remains to be elucidated. This study provides the first evidence of a proapoptotic effect and G2/M cell cycle arrest of AM251 on A375 cells. This compound may be a potential prototype for the development of promising diarylpyrazole derivatives to be evaluated in human cutaneous melanoma.
Journal/Review: ANTI-CANCER DRUGS
Volume: 26 (7) Pages from: 754 to: 762
More Information: This work was supported by the Association against Melanoma Onlus (Italy).KeyWords: A375 cells; AM251; apoptosis; Bax; Bcl-2DOI: 10.1097/CAD.0000000000000246Citations: 20data from “WEB OF SCIENCE” (of Thomson Reuters) are update at: 2024-10-13References taken from IsiWeb of Knowledge: (subscribers only)Connecting to view paper tab on IsiWeb: Click hereConnecting to view citations from IsiWeb: Click here