title: ABT-737 promotes the dislocation of ER luminal proteins to the cytosol, including pseudomonas exotoxin.
abstract: Impaired apoptosis is often a key element in tumor development. Therefore, drugs mimicking prosurvival antagonists offer promise as cancer therapeutics. When ABT-737, a BH3-only mimetic, was added to KB3-1 human cervical adenocarcinoma cells, we noted an induction of an endoplasmic reticulum (ER) stress response and the dislocation of ER luminal proteins, including chaperones, to the cell cytosol. Furthermore, when immunotoxin (antibody-toxin chimeric molecule) and ABT-737 combinations were added to cells, there was enhanced toxin-mediated inhibition of protein synthesis, consistent with enhanced translocation of toxin to the cytosol. A similar enhancement was not seen with thapsigargin, suggesting that ER stress alone was not responsible for enhanced translocation. Cytosol preparations from ABT-737-treated but not from thapsigargin-treated cells revealed the presence of greater amounts of processed 37-kDa toxin fragment compared with the addition of immunotoxin alone. As early as 4 hours after the addition of ABT-737 and immunotoxin, there was release of mitochondrial cytochrome c and activation of caspase-3/7 indicating that the combination caused apoptotic cell death. These results were reflected in decreased cellular ATP levels that were noted with combinations of ABT-737 and immunotoxin but not with either agent alone or with combinations of thapsigargin and immunotoxin. We conclude that ABT-737 increases ER permeability, promoting the dislocation of toxin from the ER to the cytosol resulting in early apoptotic cell death. These mechanistic insights suggest why this class of BH3-only mimetic synergizes in a particular way with Pseudomonas exotoxin-based immunotoxins.
authors.affiliation: Authors' Affiliation: Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland.
authors.forename: Antonella
authors.initials: A
authors.lastname: Antignani
authors.affiliation: Authors' Affiliation: Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland.
authors.forename: Robert
authors.initials: R
authors.lastname: Sarnovsky
authors.affiliation: Authors' Affiliation: Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland fitzgerd@helix.nih.gov.
authors.forename: David J
authors.initials: DJ
authors.lastname: FitzGerald
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols; D017209:Apoptosis / Q000187:drug effects; D001713:Biphenyl Compounds / Q000008:administration & dosage*; D003600:Cytosol; D004721:Endoplasmic Reticulum / Q000187:drug effects; D005098:Exotoxins / Q000008:administration & dosage* / Q000235:genetics; D005260:Female; D006801:Humans; D000922:Immunotoxins / Q000008:administration & dosage* / Q000235:genetics; D009596:Nitrophenols / Q000008:administration & dosage*; D010879:Piperazines / Q000008:administration & dosage; D014176:Protein Biosynthesis; D011549:Pseudomonas / Q000378:metabolism; D013449:Sulfonamides / Q000008:administration & dosage*; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis / Q000188:drug therapy* / Q000473:pathology
chemical_list: C501332:ABT-737; D001713:Biphenyl Compounds; D005098:Exotoxins; D000922:Immunotoxins; D009596:Nitrophenols; D010879:Piperazines; D013449:Sulfonamides
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