title: [Effect of heat shock protein 90 inhibitor 17-dimethylamino-ethylaminogeldanamycin on TNfalpha-mediated apoptosis and TNF-induced NF-kappaB activation].
abstract: OBJECTIVE
  To investigate the effect of heat shock protein 90 (HSP90) inhibitor 17-dimethylaminoethylaminogeldanamycin (17DMAG) on tumor necrosis factor alpha (TNFalpha) mediated apopotosis in cancer cells and the underlying molecular mechanisms.
  
  
  METHODS
  Cell death treated with different concentration of 17DMAG and TNFalpha was detected based on the release of lactate dehydrogenase (LDH) using a cytotoxicity detection kit. For morphological study of cell death, cells were stained with 50 microg/mL of acridine orange and 50 microg/mL of ethidium bromide and observed and photographed under a fluorescence microscope. Activation of apoptosis and NF-kappaB pathway were evaluated on the change of caspase-8, caspase-3, poly (ADP-ribose) polymerase (PARP), receptor-interaction protein (RIP), IkappaB kinase beta (IKKbeta) and inhibitor of IkappaB (IkappaBalpha) by Western blot.
  
  
  RESULTS
  17DMAG sensitized cervical cancer cells HeLa and ovarian cancer cells SKOV3 to TNFalpha-induced cell death in a dose-dependent manner, which was accompanied with degradation of RIP and Ikappakappabeta, and consequent blockage of TNFalpha-induced NFkappaB activation. 17DMAG and TNFalpha cotreated cells showed typical apoptotic morphologies and enhancing of activation of caspases.
  
  
  CONCLUSIONS
  17DMAG sensitizes cancer cells to TNFalpha-mediated apoptosis through blockage of TNF-induced NF-kappaB activation, and disabling this survival signal with 17DMAG followed by TNF treatment could be an effective new therapeutic strategy for improving the anti-cancer value of TNFalpha.
authors.affiliation: Department of Forensic Analytical Toxicology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
authors.forename: Ying-Jia
authors.initials: YJ
authors.lastname: Zhong
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.forename: Xue-Lian
authors.initials: XL
authors.lastname: Zheng
authors.forename: You-Yi
authors.initials: YY
authors.lastname: Yan
authors.forename: Xiao-Min
authors.initials: XM
authors.lastname: Liu
authors.forename: Lin-Chuan
authors.initials: LC
authors.lastname: Liao
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D016227:Benzoquinones / Q000494:pharmacology*; D045744:Cell Line, Tumor; D005260:Female; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors*; D006367:HeLa Cells; D006801:Humans; D047029:Lactams, Macrocyclic / Q000494:pharmacology*; D016328:NF-kappa B / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000473:pathology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; C112765:tanespimycin