title: Cell death induction by the BH3 mimetic GX15-070 in thyroid carcinoma cells.
abstract: BACKGROUND
  The evasion of cell death is one of the hallmarks of cancer, contributing to both tumor progression and resistance to therapy. Dedifferentiated and anaplastic thyroid carcinomas that do not take up radioiodine are resistant to conventional anticancer treatments and patients with these tumors are difficult to treat. BH3 mimetics are a new class of drugs that target anti-apoptotic proteins of the BCL-2 family and promote cell death. The purpose of this study was to analyze the molecular effects of the BH3 mimetic GX15-070 on thyroid carcinoma cell lines and to characterize cell death induced by GX15-070.
  
  
  METHODS
  A total of 17 cell lines derived from follicular, papillary, and anaplastic thyroid carcinomas were treated with GX15-070. Cell viability was measured with MTT assay while cell cycle phase distribution and subG1 peaks were determined after propidium iodide staining. We assessed cell death via the caspase 3/7 activity, caspase cleavage products, lactate dehydrogenase (LDH) liberation assays, and a LC3 analysis by western blot. Ultrastructural changes were analysed by electron microscopy of GX15-070-treated cells.
  
  
  RESULTS
  After GX15-070 treatment, the number of viable cells was decreased in all cell lines examined, with IC50 values ranging from 48nM to 3.25 μM. We observed biochemical markers of autophagic cell death and necrosis like LC3 conversion and LDH release after the GX15-070 treatment. Electron microscopy revealed several common characteristic ultrastructural changes like swelling of mitochondria, dilatation of rough endoplasmic reticulum, membrane blebbing and formation of vacuoles. GX15-070 treatment induced DNA fragmentation detected by subG1-peak induction and an arrest in G1 phase of the cell cycle. Caspase activation after GX15-070 incubation was detected but had no effect on viability of cells.
  
  
  CONCLUSIONS
  With these experiments we demonstrated the efficacy of the BH3 mimetic drug GX15-070 acting against dedifferentiated thyroid carcinoma cells of various histological origins by the induction of cell death. GX15-070-treated cells underwent non-classical cell death with signs of apoptosis, autophagy and necrosis in parallel. GX15-07 and related compounds thus may be a new therapeutic option for dedifferentiated thyroid carcinoma of various histological subtypes.
authors.affiliation: Department of Endocrinology and Metabolism, and Division of Laboratory Research, University Hospital Essen, Hufelandstrasse 55, Essen, Germany. martina.broecker@uni-due.de.|Present address: Department of Clinical Chemistry, University Hospital Essen, Hufelandstrasse 55, 45122, Essen, Germany. martina.broecker@uni-due.de.
authors.forename: Martina
authors.initials: M
authors.lastname: Broecker-Preuss
authors.affiliation: Department of Endocrinology and Metabolism, and Division of Laboratory Research, University Hospital Essen, Hufelandstrasse 55, Essen, Germany. naj@gmx.de.|Present address: Ruhrlandklinik, University Hospital Essen, Tüschener Weg 40, 45239, Essen, Germany. naj@gmx.de.
authors.forename: Jan
authors.initials: J
authors.lastname: Viehof
authors.affiliation: Institute of Anatomy, University Hospital Essen, Hufelandstrasse 55, Essen, Germany. holger.jastrow@uk-essen.de.
authors.forename: Holger
authors.initials: H
authors.lastname: Jastrow
authors.affiliation: Department of Endocrinology and Metabolism, and Division of Laboratory Research, University Hospital Essen, Hufelandstrasse 55, Essen, Germany. nina.becher-boveleth@uk-essen.de.|Present address: Clinic of Nuclear Medicine, University Hospital Essen, Hufelandstr. 55, 45122, Essen, Germany. nina.becher-boveleth@uk-essen.de.
authors.forename: Nina
authors.initials: N
authors.lastname: Becher-Boveleth
authors.affiliation: Department of Endocrinology and Metabolism, and Division of Laboratory Research, University Hospital Essen, Hufelandstrasse 55, Essen, Germany. dagmar.fuehrer-sakel@uk-essen.de.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Fuhrer
authors.affiliation: Department of Endocrinology and Metabolism, and Division of Laboratory Research, University Hospital Essen, Hufelandstrasse 55, Essen, Germany. klaus.mann@ezah.de.|Present address: Center of Endocrinology Alter Hof München, Dienerstr. 12, 80331, Munich, Germany. klaus.mann@ezah.de.
authors.forename: Klaus
authors.initials: K
authors.lastname: Mann
mesh_terms: D017209:Apoptosis; D016923:Cell Death; D045744:Cell Line, Tumor; D002470:Cell Survival; D006801:Humans; D007211:Indoles; D058990:Molecular Targeted Therapy / Q000379:methods*; D011758:Pyrroles / Q000008:administration & dosage / Q000627:therapeutic use*; D013964:Thyroid Neoplasms / Q000188:drug therapy*
chemical_list: D007211:Indoles; D011758:Pyrroles; C520962:obatoclax