title: Cytotoxic effect of disulfiram/copper on human glioblastoma cell lines and ALDH-positive cancer-stem-like cells.
abstract: BACKGROUND
  Glioblastoma multiforme (GBM) cells are resistant to anticancer drugs. Cancer stem cells (CSCs) are a key mediator of chemoresistance. We have reported that disulfiram (DS), an aldehyde dehydrogenase (ALDH) inhibitor, targets breast CSC-like cells. In this study, the effect of DS and combination of DS and gemcitabine (dFdC) on GBM cells and GBM stem-like cells was investigated.
  
  
  METHODS
  1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), combination index (CI)-isobologram, western blot, luciferase reporter gene assay, electrophoretic mobility-shift assay and ALDH analysis were used in this study.
  
  
  RESULTS
  Disulfiram is cytotoxic in GBM cell lines in a copper (Cu)-dependent manner. Disulfiram/copper enhances the cytotoxicity of dFdC. Combination index-isobologram analysis indicates a synergistic effect between DS/Cu and dFdC. Disulfiram/copper induces reactive oxygen species (ROS), activates JNK and p38 pathways and inhibits nuclear factor-kappa B activity in GBM cell lines. Disulfiram/copper may trigger intrinsic apoptotic pathway via modulation of the Bcl2 family. Disulfiram/copper abolishes stem-like cell population in GBM cell lines.
  
  
  CONCLUSIONS
  Our findings indicate that the cytotoxicity of DS/Cu and the enhancing effect of DS/Cu on the cytotoxicity of dFdC in GBM stem-like cells may be caused by induction of ROS and inhibition of both ALDH and the NFkB pathway. Both DS and dFdC can traverse the blood-brain barrier. Further study may lead them into GBM chemotherapy.
authors.affiliation: Research Institute in Healthcare Science, School of Applied Sciences, University of Wolverhampton, Wolverhampton, UK.
authors.forename: P
authors.initials: P
authors.lastname: Liu
authors.forename: S
authors.initials: S
authors.lastname: Brown
authors.forename: T
authors.initials: T
authors.lastname: Goktug
authors.forename: P
authors.initials: P
authors.lastname: Channathodiyil
authors.forename: V
authors.initials: V
authors.lastname: Kannappan
authors.forename: J-P
authors.initials: JP
authors.lastname: Hugnot
authors.forename: P-O
authors.initials: PO
authors.lastname: Guichet
authors.forename: X
authors.initials: X
authors.lastname: Bian
authors.forename: A L
authors.initials: AL
authors.lastname: Armesilla
authors.forename: J L
authors.initials: JL
authors.lastname: Darling
authors.forename: W
authors.initials: W
authors.lastname: Wang
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects; D001932:Brain Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D003300:Copper / Q000494:pharmacology*; D003602:Cytotoxicity, Immunologic; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D004221:Disulfiram / Q000494:pharmacology*; D004357:Drug Synergism; D005909:Glioblastoma / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D006801:Humans; D048670:MAP Kinase Kinase 4 / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D017382:Reactive Oxygen Species / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D000093542:Gemcitabine
chemical_list: D016328:NF-kappa B; D017382:Reactive Oxygen Species; D003841:Deoxycytidine; D003300:Copper; D000444:Aldehyde Dehydrogenase; D048051:p38 Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4; D004221:Disulfiram; D000093542:Gemcitabine