title: Protein kinase C inhibitors sensitize GNAQ mutant uveal melanoma cells to ionizing radiation.
abstract: OBJECTIVE
  Uveal melanoma (UM) tumors require large doses of radiation therapy (RT) to achieve tumor ablation, which frequently results in damage to adjacent normal tissues, leading to vision-threatening complications. Approximately 50% of UM patients present with activating somatic mutations in the gene encoding for G protein αq-subunit (GNAQ), which lead to constitutive activation of downstream pathways, including protein kinase C (PKC). In this study, we investigated the impact of small-molecule PKC inhibitors bisindolylmaleimide I (BIM) and sotrastaurin (AEB071), combined with ionizing radiation (IR), on survival in melanoma cell lines.
  
  
  METHODS
  Cellular radiosensitivity was determined by using a combination of proliferation, viability, and clonogenic assays. Cell-cycle effects were measured by flow cytometry. Transcriptomic and proteomic profiling were performed by quantitative real-time PCR, reverse-phase protein array analysis, and immunofluorescence.
  
  
  RESULTS
  We found that the PKC inhibitors combined with IR significantly decreased the viability, proliferation, and clonogenic potential of GNAQ(mt), but not GNAQ(wt)/BRAF(mt) cells, compared with IR alone. Combined treatment increased the antiproliferative and proapoptotic effects of IR in GNAQ(mt) cells through delayed DNA-damage resolution and enhanced induction of proteins involved in cell-cycle arrest, cell-growth arrest, and apoptosis.
  
  
  CONCLUSIONS
  Our preclinical results suggest that combined modality treatment may allow for reductions in the total RT dose and/or fraction size, which may lead to better functional organ preservation in the treatment of primary GNAQ(mt) UM. These findings suggest future clinical trials combining PKC inhibitors with RT in GNAQ(mt) UM warrant consideration.
authors.affiliation: Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, United States.
authors.forename: Jasmina Ziva
authors.initials: JZ
authors.lastname: Cerne
authors.forename: Sean Michael
authors.initials: SM
authors.lastname: Hartig
authors.forename: Mark Patrick
authors.initials: MP
authors.lastname: Hamilton
authors.forename: Sue Anne
authors.initials: SA
authors.lastname: Chew
authors.forename: Nicholas
authors.initials: N
authors.lastname: Mitsiades
authors.forename: Vassiliki
authors.initials: V
authors.lastname: Poulaki
authors.forename: Sean Eric
authors.initials: SE
authors.lastname: McGuire
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D002453:Cell Cycle / Q000187:drug effects / Q000528:radiation effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D003131:Combined Modality Therapy; D004273:DNA, Neoplasm / Q000235:genetics*; D005434:Flow Cytometry; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D006801:Humans; D008545:Melanoma / Q000201:enzymology* / Q000235:genetics / Q000628:therapy; D009154:Mutation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011839:Radiation, Ionizing; D060888:Real-Time Polymerase Chain Reaction; D014604:Uveal Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000628:therapy; D000098943:Uveal Melanoma
chemical_list: D004273:DNA, Neoplasm; C554061:GNAQ protein, human; D044385:GTP-Binding Protein alpha Subunits; D047428:Protein Kinase Inhibitors; D043802:GTP-Binding Protein alpha Subunits, Gq-G11
keywords: GNAQ mutation; protein kinase C; radiation therapy; radiosensitization; uveal melanoma