title: Anti-tumour activity of fotemustine and protons in combination with bevacizumab.
abstract: BACKGROUND
  Metastatic melanoma is one of the most aggressive tumours and is also very resistant to current therapeutic approaches. The aim of this investigation was the in vitro study of the anti-proliferative effects of fotemustine (FM; 100 and 250 microM), bevacizumab (5 microg/ml) and proton irradiation (12 and 16 Gy) on resistant HTB140 human melanoma cells.
  
  
  METHODS
  Viability was estimated by sulphorhodamine B assay, while cell proliferation was analyzed by 5-bromo-2-deoxyuridine assay. Cell cycle distribution and apoptosis were examined using flow cytometry.
  
  
  RESULTS
  Cell viability and proliferation were reduced after all applied treatments. The level of apoptosis significantly increased after treatment with FM, protons or a combination of all agents, while the apoptotic index ranged from 1.2 to 9.2. Proton irradiation, as well as combined treatment with bevacizumab and protons or 100 microM FM, bevacizumab and protons, have reduced melanoma cell proliferation through the induction of G1 phase arrest. Single FM (250 microM) or bevacizumab treatment and their combination, as well as the joint application of these 2 agents with protons, reduced cell proliferation and provoked G2 phase accumulation.
  
  
  CONCLUSIONS
  The analyzed treatments reduced cell viability and proliferation, triggered G1 or G2 cell cycle phase accumulation and stimulated apoptotic cell death.
authors.affiliation: Vinca Institute of Nuclear Sciences, Belgrade, Serbia.
authors.forename: Lela B
authors.initials: LB
authors.lastname: Korićanac
authors.forename: Jelena J
authors.initials: JJ
authors.lastname: Zakula
authors.forename: Ivan M
authors.initials: IM
authors.lastname: Petrović
authors.forename: Lucia M
authors.initials: LM
authors.lastname: Valastro
authors.forename: Giuseppe A P
authors.initials: GA
authors.lastname: Cirrone
authors.forename: Giacomo
authors.initials: G
authors.lastname: Cuttone
authors.forename: Aleksandra M
authors.initials: AM
authors.lastname: Ristić-Fira
mesh_terms: D000911:Antibodies, Monoclonal / Q000008:administration & dosage*; D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000008:administration & dosage; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D000068258:Bevacizumab; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D006801:Humans; D009607:Nitrosourea Compounds / Q000008:administration & dosage*; D009943:Organophosphorus Compounds / Q000008:administration & dosage*; D011522:Protons
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents; D009607:Nitrosourea Compounds; D009943:Organophosphorus Compounds; D011522:Protons; D000068258:Bevacizumab; C054368:fotemustine