title: AurkA inhibitors enhance the effects of B-RAF and MEK inhibitors in melanoma treatment.
abstract: BACKGROUND
  Aurora kinase A (AurkA) is over-expressed in melanoma and its inhibition has been observed to limit tumor growth, suggesting a potential role in melanoma treatment.
  
  
  METHODS
  A human melanoma cell line with the B-RAF (V600E) mutation (A375mel) was exposed to B-RAF inhibitor (GSK2118436), MEK inhibitor (GSK1120212) and AurkA inhibitor (MLN8054) as single agents or in various combinations (BRAF plus AurkA inhibitor, MEK plus AurkA inhibitor or triple combination BRAF plus MEK plus AurkA inhibitor). Cell proliferation was assessed using xCELLigence technology. Total protein extracts were examined for p53 and c-Myc protein expression by Western blot analysis. Drug anti-tumor effects were further assessed using a 3D-human melanoma skin reconstruction model, in which tissues were incubated with serum-free medium containing control, B-RAF plus MEK inhibitor, MEK plus AurkA inhibitor or the triple combination.
  
  
  RESULTS
  AurkA inhibitor plus B-RAF inhibitor, AurkA inhibitor plus MEK inhibitor or triple combination had a markedly greater anti-proliferative effect on A375 (BRAFV600E) melanoma cells than single agents. In the 3D human skin model, the triple combination had a greater anti-tumor effect at the epidermal/dermal junction than control or either double combination. However, S-100 and Ki-67 positively stained spindle-shaped cells were detected in the dermal stratum, suggesting the presence of alive and proliferating melanoma cells.
  
  
  CONCLUSIONS
  These findings provide new prospects for melanoma research, including combined B-RAF/AurkA inhibition for B-RAF mutated melanomas and MEK/AurkA inhibitor combination for patients without B-RAF mutations. Moreover, for the first time, we have shown that a B-RAF, MEK and AurkA inhibitor triple drug combination offers increased efficacy against melanoma cell growth and might be considered as a potential treatment strategy for enhancing clinical response in melanoma. However, although this triple drug combination was more effective at the epidermal/dermal junction, the suggested presence of alive and proliferating melanoma cells in the dermal stratum could result in drug resistance and disease recurrence. Molecular characterization of these dermal cells may be critical for the development of novel therapeutic strategies.
authors: Emilia
authors: E
authors: Caputo
authors: Roberta
authors: R
authors: Miceli
authors: Maria Letizia
authors: ML
authors: Motti
authors: Rosarita
authors: R
authors: Taté
authors: Federica
authors: F
authors: Fratangelo
authors: Gerardo
authors: G
authors: Botti
authors: Nicola
authors: N
authors: Mozzillo
authors: Maria Vincenza
authors: MV
authors: Carriero
authors: Ernesta
authors: E
authors: Cavalcanti
authors: Giuseppe
authors: G
authors: Palmieri
authors: Gennaro
authors: G
authors: Ciliberto
authors: Giuseppe
authors: G
authors: Pirozzi
authors: Paolo Antonio
authors: PA
authors: Ascierto
mesh_terms: D064096:Aurora Kinase A / Q000037:antagonists & inhibitors* / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000378:metabolism; D008545:Melanoma / Q000188:drug therapy* / Q000201:enzymology*; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008954:Models, Biological; D047428:Protein Kinase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use*; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012867:Skin / Q000187:drug effects
chemical_list: D019394:Ki-67 Antigen; D047428:Protein Kinase Inhibitors; D064096:Aurora Kinase A; D048493:Proto-Oncogene Proteins B-raf; D020929:Mitogen-Activated Protein Kinase Kinases