title: Endonucleases induced TRAIL-insensitive apoptosis in ovarian carcinoma cells.
abstract: TRAIL induced apoptosis of tumor cells is currently entering phase II clinical settings, despite the fact that not all tumor types are sensitive to TRAIL. TRAIL resistance in ovarian carcinomas can be caused by a blockade upstream of the caspase 3 signaling cascade. We explored the ability of restriction endonucleases to directly digest DNA in vivo, thereby circumventing the caspase cascade. For this purpose, we delivered enzymatically active endonucleases via the cationic amphiphilic lipid SAINT-18((R)):DOPE to both TRAIL-sensitive and insensitive ovarian carcinoma cells (OVCAR and SKOV-3, respectively). Functional nuclear localization after delivery of various endonucleases (BfiI, PvuII and NucA) was indicated by confocal microscopy and genomic cleavage analysis. For PvuII, analysis of mitochondrial damage demonstrated extensive apoptosis both in SKOV-3 and OVCAR. This study clearly demonstrates that cellular delivery of restriction endonucleases holds promise to serve as a novel therapeutic tool for the treatment of resistant ovarian carcinomas.
authors.affiliation: Department of Pathology and Medical Biology, Groningen University Institute for Drug Exploration, University Medical Center Groningen, Hanzeplein 1, Groningen, The Netherlands.
authors.forename: Tessa M
authors.initials: TM
authors.lastname: Geel
authors.forename: Gregor
authors.initials: G
authors.lastname: Meiss
authors.forename: Bernardina T
authors.initials: BT
authors.lastname: van der Gun
authors.forename: Bart Jan
authors.initials: BJ
authors.lastname: Kroesen
authors.forename: Lou F
authors.initials: LF
authors.lastname: de Leij
authors.forename: Mindaugas
authors.initials: M
authors.lastname: Zaremba
authors.forename: Arūnas
authors.initials: A
authors.lastname: Silanskas
authors.forename: Michael
authors.initials: M
authors.lastname: Kokkinidis
authors.forename: Alfred
authors.initials: A
authors.lastname: Pingoud
authors.forename: Marcel H
authors.initials: MH
authors.lastname: Ruiters
authors.forename: Pamela M
authors.initials: PM
authors.lastname: McLaughlin
authors.forename: Marianne G
authors.initials: MG
authors.lastname: Rots
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D053938:DNA Fragmentation; D004720:Endonucleases / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006657:Histones / Q000378:metabolism; D006801:Humans; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology; D010714:Phosphatidylethanolamines / Q000378:metabolism; D011726:Pyridinium Compounds / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics / Q000378:metabolism*
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; C532953:1-methyl-4-(9-dioleyl)methylpyridinium; D000590:Amino Acid Chloromethyl Ketones; D004791:Enzyme Inhibitors; C468783:H2AX protein, human; D006657:Histones; D010714:Phosphatidylethanolamines; D011726:Pyridinium Compounds; D053221:TNF-Related Apoptosis-Inducing Ligand; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D004720:Endonucleases; D053148:Caspase 3