title: The response of human ectomesenchymal dental pulp stem cells to cisplatin treatment.
abstract: OBJECTIVE
  To determine the response of dental pulp stem cells (DPSCs) to DNA-damaging cytostatic cisplatin and compare it with the response of normal human dermal fibroblasts (HDFs).
  
  
  METHODS
  Dental pulp stem cells were exposed to 5, 10, 20 or 40 μmol L(-1) of cisplatin. The proliferation of affected cells was assessed by a Z2 Counter and viability was assessed by means of a Vi-Cell XR using Trypan blue exclusion staining. Cell cycle analysis and induction of apoptosis were performed by flow cytometry. Induction of apoptosis was determined by monitoring the activities of caspases. The expression of proteins was detected by electrophoresis and Western blotting. The descriptive statistics of the results was analyzed by Student's t-test.
  
  
  RESULTS
  Dental pulp stem cells had a greater genotoxic stress response to cisplatin compared to HDFs. All three main Mitogen-activated protein kinases (MAPK) families - extracellular signal-regulated kinases (ERK), c-Jun-N-terminal kinase (JNK) and p38 were activated after treatment of DPSCs with cisplatin. The activation of MAPK pathways was not observed in HDFs exposed to cisplatin. The exposure of DPSCs and HDFs to cisplatin provoked an increase in p53 and p21 expression and p53 phosphorylation of serine 15. Higher concentrations of cisplatin reduced the viability of DPSCs and HDFs and induced the activation of caspases 3/7 and 9.
  
  
  CONCLUSIONS
  Dental pulp stem cells had a greater genotoxic stress response to cisplatin compared to HDFs. Cisplatin in higher concentrations triggered activation of MAPK and apoptosis in DPSCs but not in HDFs.
authors.affiliation: Departments of Medical Biochemistry Histology and Embryology, Faculty of Medicine in Hradec Králové, Charles University in Prague, Czech Republic. seifrtovam@lfhk.cuni.cz
authors.forename: M
authors.initials: M
authors.lastname: Seifrtová
authors.forename: R
authors.initials: R
authors.lastname: Havelek
authors.forename: J
authors.initials: J
authors.lastname: Cmielová
authors.forename: A
authors.initials: A
authors.lastname: Jiroutová
authors.forename: T
authors.initials: T
authors.lastname: Soukup
authors.forename: L
authors.initials: L
authors.lastname: Brůčková
authors.forename: J
authors.initials: J
authors.lastname: Mokrý
authors.forename: D
authors.initials: D
authors.lastname: English
authors.forename: M
authors.initials: M
authors.lastname: Rezáčová
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000187:drug effects; D053179:Caspase 7 / Q000187:drug effects; D053181:Caspase 8 / Q000187:drug effects; D053453:Caspase 9 / Q000187:drug effects; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000633:toxicity*; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000187:drug effects; D054697:Cytostatic Agents / Q000633:toxicity*; D003782:Dental Pulp / Q000166:cytology* / Q000187:drug effects; D004475:Ectoderm / Q000166:cytology* / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000187:drug effects; D020935:MAP Kinase Signaling System / Q000187:drug effects; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D009153:Mutagens / Q000633:toxicity; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D012694:Serine / Q000187:drug effects; D012867:Skin / Q000166:cytology / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects; D048051:p38 Mitogen-Activated Protein Kinases / Q000187:drug effects
chemical_list: C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D054697:Cytostatic Agents; D009153:Mutagens; D047428:Protein Kinase Inhibitors; D016159:Tumor Suppressor Protein p53; D012694:Serine; D048049:Extracellular Signal-Regulated MAP Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C505530:CASP8 protein, human; D053148:Caspase 3; D053179:Caspase 7; D053181:Caspase 8; D053453:Caspase 9; D002945:Cisplatin