title: Analysis of the Antitumor Activity of Clotrimazole on A375 Human Melanoma Cells.
abstract: OBJECTIVE
  The current study was designed to characterize the anticancer effects of clotrimazole on human cutaneous melanoma cells.
  
  
  METHODS
  The v-raf murine sarcoma viral oncogene homolog B1 V600E mutant melanoma cell line A375 was used as an in vitro model. Characterization tools included analyses of cell viability, gene expression, cell-cycle progression, annexin V reactivity and internucleosomal DNA fragmentation.
  
  
  RESULTS
  Clotrimazole induced cytotoxicity in A375 human melanoma cells without significant changes of human keratinocyte cell viability. Clotrimazole, at a concentration that approximates the inhibitory concentration 50% (IC50) value (i.e. 10 μM), reduced the expression of hexokinase type-II, induced cell-cycle arrest at G1-S phase transition, altered annexin V reactivity and induced DNA fragmentation without evidence of necrosis.
  
  
  CONCLUSIONS
  The current study provides evidence of a remarkable pro-apoptotic effect by clotrimazole against human melanoma cells, with a different mechanism of action and timeline of the apoptosis-related events when compared to cisplatin.
authors.affiliation: Nello Carrara Institute of Applied Physics, National Research Council, Sesto Fiorentino, Florence, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Adinolfi
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Carpi
authors.affiliation: Medical Oncology Unit, University Hospital of Pisa, Pisa, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Romanini
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Da Pozzo
authors.affiliation: Department of Translational Research and of New Surgical and Medical Technologies, University of Pisa, Pisa, Italy.
authors.forename: Maura
authors.initials: M
authors.lastname: Castagna
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Costa
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Martini
authors.affiliation: Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Søren-Peter
authors.initials: SP
authors.lastname: Olesen
authors.affiliation: Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Nicole
authors.initials: N
authors.lastname: Schmitt
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Breschi
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Nieri
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy stefano.fogli@farm.unipi.it.
authors.forename: Stefano
authors.initials: S
authors.lastname: Fogli
mesh_terms: D017304:Annexin A5 / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D003022:Clotrimazole / Q000494:pharmacology*; D053938:DNA Fragmentation / Q000187:drug effects; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism
chemical_list: D017304:Annexin A5; D000970:Antineoplastic Agents; D003022:Clotrimazole; D002945:Cisplatin
keywords: A375 cells; Clotrimazole; apoptosis; cell cycle; hexokinase; melanoma