title: Comparison of the cellular molecular stress responses after treatments used in bladder cancer.
abstract: OBJECTIVE
  To investigate the molecular stress responses related to the quality of recovery of normal tissue after various treatments for bladder cancer, i.e. hyperthermia, ionizing radiation, mitomycin-C and 5-aminolaevulinic acid photodynamic therapy (ALA-PDT).
  
  
  METHODS
  The study focused particularly on intracellular fibroblast levels of heat-shock protein-47 (HSP47) and HSP72, which are associated with collagen metabolism and the development of tolerance to repeated treatment, respectively. Iso-effective treatment doses (50% clonogenic cell survival) of each method were delivered to a 3T6 murine fibroblast model. Intracellular extracts were analysed at 3, 6, 9, 12 and 24 h after treatment, using Western blot analysis to compare the levels of HSP47 and HSP72. Time-matched treatment and control groups were quantified by comparison with actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression using appropriate software.
  
  
  RESULTS
  There were various changes in levels of HSP expression with treatment method; HSP47 levels were significantly higher after hyperthermia and radiation but not with mitomycin-C or ALA-PDT. HSP72 levels were significantly higher with all methods except ALA-PDT.
  
  
  CONCLUSIONS
  Hyperthermia and ionizing radiation are associated with early increases in levels of HSP47 (a marker of collagen metabolism), in contrast to ALA-PDT and mitomycin-C. These findings are compatible with clinical findings where fibrosis/scarring is common with the first two but not the last two methods. In addition, all methods except ALA-PDT are associated with an increase in HSP 72 (a protein associated with cellular tolerance) and this may help to explain, at a cellular level, why resistance to repeated ALA-PDT treatments does not seem to occur.
authors.affiliation: CRC Experimental Radiation Oncology Group, Paterson Institute for Cancer Research, Manchester, UK. Daveshackley@hotmail.com
authors.forename: D C
authors.initials: DC
authors.lastname: Shackley
authors.forename: A
authors.initials: A
authors.lastname: Haylett
authors.forename: C
authors.initials: C
authors.lastname: Whitehurst
authors.forename: C D
authors.initials: CD
authors.lastname: Betts
authors.forename: K
authors.initials: K
authors.lastname: O'Flynn
authors.forename: N W
authors.initials: NW
authors.lastname: Clarke
authors.forename: J V
authors.initials: JV
authors.lastname: Moore
mesh_terms: D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000627:therapeutic use; D015153:Blotting, Western; D003094:Collagen / Q000378:metabolism*; D050958:HSP47 Heat-Shock Proteins; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006979:Hyperthermia, Induced / Q000379:methods; D051379:Mice; D016685:Mitomycin / Q000627:therapeutic use; D010778:Photochemotherapy / Q000379:methods; D013312:Stress, Physiological / Q000209:etiology / Q000378:metabolism; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000628:therapy*
chemical_list: D000903:Antibiotics, Antineoplastic; D050958:HSP47 Heat-Shock Proteins; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins; C494661:Serpinh1 protein, mouse; D016685:Mitomycin; D003094:Collagen