title: Restoration by glycerol of p53-dependent apoptosis in cells bearing the mutant p53 gene.
abstract: OBJECTIVE
  Effective heat-induced cell death in cultured cells bearing a mutant p53 (mp53) gene was sought by glycerol treatment which led to conformational change from mp53 to wild-type p53 (wtp53) in p53-null murine fibroblasts transfected with mp53.
  
  
  METHODS
  Heat sensitivity was measured using a colony-forming assay. For heat-induced apoptosis, gel electrophoresis was applied to detect DNA fragmentation and Hoechst33342 staining for apoptotic bodies. Glycerol (0.6 M) was applied to the cultured cells 48 h before heating at 44 degrees C in a water bath.
  
  
  RESULTS
  Wtp53 transfectants (MT158/wtp53-1) were sensitive to heat stress compared with mp53 transfectants (MT158/mp53-2 cells), and the combined treatment with glycerol enhanced cell killing only in the MT158/mp53-2 cells. After glycerol pretreatment for 48 h, the subsequent heat treatment enhanced DNA fragmentation and apoptosis in MT158/mp53-2 cells, while DNA fragmentation and apoptotic bodies were not enhanced with heat treatment alone in these cells. In contrast, DNA fragmentation or apoptotic bodies were clearly observed in MT158/wtp53-1 cells 3-24h after heat treatment. Treatment with glycerol alone did not induce apoptosis in the transfectants.
  
  
  CONCLUSIONS
  Glycerol appears to function as a chemical chaperone that restores mp53 to wtp53 function.
authors.affiliation: Department of Biology, Nara Medical University, Kashihara, Japan. tohnishi@nmu-gw.naramed-u.ac.jp
authors.forename: T
authors.initials: T
authors.lastname: Ohnishi
authors.forename: H
authors.initials: H
authors.lastname: Matsumoto
authors.forename: X
authors.initials: X
authors.lastname: Wang
authors.forename: A
authors.initials: A
authors.lastname: Takahashi
authors.forename: T
authors.initials: T
authors.lastname: Tamamoto
authors.forename: K
authors.initials: K
authors.lastname: Ohnishi
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D053938:DNA Fragmentation / Q000187:drug effects; D016158:Genes, p53 / Q000502:physiology*; D005990:Glycerol / Q000494:pharmacology*; D051379:Mice; D009154:Mutation
chemical_list: D005990:Glycerol