title: Combined inhibition of the cell cycle related proteins Wee1 and Chk1/2 induces synergistic anti-cancer effect in melanoma.
abstract: BACKGROUND
  Malignant melanoma has an increasing incidence rate and the metastatic disease is notoriously resistant to standard chemotherapy. Loss of cell cycle checkpoints is frequently found in many cancer types and makes the cells reliant on compensatory mechanisms to control progression. This feature may be exploited in therapy, and kinases involved in checkpoint regulation, such as Wee1 and Chk1/2, have thus become attractive therapeutic targets.
  
  
  METHODS
  In the present study we combined a Wee1 inhibitor (MK1775) with Chk1/2 inhibitor (AZD7762) in malignant melanoma cell lines grown in vitro (2D and 3D cultures) and in xenografts models.
  
  
  RESULTS
  Our in vitro studies showed that combined inhibition of Wee1 and Chk1/2 synergistically decreased viability and increased apoptosis (cleavage of caspase 3 and PARP), which may be explained by accumulation of DNA-damage (increased expression of γ-H2A.X)--and premature mitosis of S-phase cells. Compared to either inhibitor used as single agents, combined treatment reduced spheroid growth and led to greater tumour growth inhibition in melanoma xenografts.
  
  
  CONCLUSIONS
  These data provide a rationale for further evaluation of the combination of Wee1 and Chk1/2 inhibitors in malignant melanoma.
authors.affiliation: Department of Pathology, Oslo University Hospital, Norwegian Radium Hospital, N-0310, Oslo, Norway. Gry.Magnussen@rr-research.no.
authors.forename: Gry Irene
authors.initials: GI
authors.lastname: Magnussen
authors.affiliation: Department of Pathology, Oslo University Hospital, Norwegian Radium Hospital, N-0310, Oslo, Norway. Elisabeth.Emilsen@rr-research.no.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Emilsen
authors.affiliation: Department of Tumour Biology, Institute for Cancer Research, Norwegian Radium Hospital, Oslo, Norway. Karianne.Giller.Fleten@rr-research.no.
authors.forename: Karianne
authors.initials: K
authors.lastname: Giller Fleten
authors.affiliation: Department of Tumour Biology, Institute for Cancer Research, Norwegian Radium Hospital, Oslo, Norway. Birgit.Engesater@rr-research.no.
authors.forename: Birgit
authors.initials: B
authors.lastname: Engesæter
authors.affiliation: Department of Radiation Biology, Institute for Cancer Research, Norwegian Radium Hospital, Oslo, Norway. Viola.Naehse-Kumpf@curie.fr.
authors.forename: Viola
authors.initials: V
authors.lastname: Nähse-Kumpf
authors.affiliation: Department of Medical Genetics, Ullevål University Hospital, Oslo, Norway. Roar.Fjar@medisin.uio.no.
authors.forename: Roar
authors.initials: R
authors.lastname: Fjær
authors.affiliation: Department of Pathology, Oslo University Hospital, Norwegian Radium Hospital, N-0310, Oslo, Norway. Ana.Slipicevic@rr-research.no.
authors.forename: Ana
authors.initials: A
authors.lastname: Slipicevic
authors.affiliation: Department of Pathology, Oslo University Hospital, Norwegian Radium Hospital, N-0310, Oslo, Norway. vivi.ann.florenes@radiumhospitalet.no.
authors.forename: Vivi Ann
authors.initials: VA
authors.lastname: Flørenes
mesh_terms: D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000008:administration & dosage; D017209:Apoptosis / Q000187:drug effects; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D064447:Checkpoint Kinase 2 / Q000037:antagonists & inhibitors / Q000235:genetics*; D004357:Drug Synergism; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D051379:Mice; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics*; D011720:Pyrazoles / Q000008:administration & dosage; D011743:Pyrimidines / Q000008:administration & dosage; D011744:Pyrimidinones; D012878:Skin Neoplasms; D013876:Thiophenes / Q000008:administration & dosage; D014508:Urea / Q000008:administration & dosage / Q000031:analogs & derivatives; D023041:Xenograft Model Antitumor Assays; D000096142:Cutaneous Malignant Melanoma
chemical_list: C532363:3-(carbamoylamino)-5-(3-fluorophenyl)-N-(3-piperidyl)thiophene-2-carboxamide; D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011743:Pyrimidines; D011744:Pyrimidinones; D013876:Thiophenes; D014508:Urea; D064447:Checkpoint Kinase 2; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; C578634:CHEK2 protein, human; C549567:adavosertib