title: N(alpha)-tosyl-L-phenylalanine chloromethyl ketone induces caspase-dependent apoptosis in transformed human B cell lines with transcriptional down-regulation of anti-apoptotic HS1-associated protein X-1.
abstract: N(alpha)-tosyl-L-phenylalanine chloromethylketone (TPCK) has been widely used to investigate signal transduction pathways that are involved in gene expression and cell survival/cell death. However, contradictory effects of TPCK on apoptosis have been reported, and the underlying signaling events leading to TPCK-induced promotion or prevention of apoptosis are not fully understood. Here, we show that TPCK induces caspase-dependent apoptosis in Epstein-Barr virus (EBV)-transformed human B cell lines with release of pro-apoptotic proteins from mitochondria. TPCK treatment also results in down-regulation of the anti-apoptotic proteins, cIAP1, cIAP2, and HAX-1, and caspase-dependent cleavage of the anti-apoptotic proteins, Bcl-2 and XIAP. Quantitative PCR analysis confirmed that the TPCK-induced down-regulation of HAX-1 occurred at the transcriptional level, and experiments using the specific pharmacological inhibitor, Bay 11-7082, suggested that HAX-1 expression is subject to regulation by the transcription factor, NF-kappaB. B cell lines derived from patients with homozygous HAX1 mutations were more sensitive to TPCK-induced apoptosis when compared with normal donor cell lines. Furthermore, N-acetylcysteine effectively blocked TPCK-induced apoptosis in EBV-transformed B cell lines and prevented the down-regulation or cleavage of anti-apoptotic proteins. Taken together, our studies demonstrate that TPCK induces apoptosis in human B cell lines and exerts multiple effects on pro- and anti-apoptotic factors.
authors.affiliation: Division of Molecular Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm 171 76, Sweden; Department of Biochemistry, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Thalassemia Research Center, Institute of Science and Technology for Research and Development, Mahidol University, Nakhonpathom 73170, Thailand.
authors.forename: Siriporn
authors.initials: S
authors.lastname: Jitkaew
authors.affiliation: Department of Molecular Biology, The Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw 02-781, Poland.
authors.forename: Alicja
authors.initials: A
authors.lastname: Trebinska
authors.affiliation: Department of Molecular Biology, The Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw 02-781, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Grzybowska
authors.affiliation: Childhood Cancer Research Unit, Department of Woman and Child Health, Karolinska Institutet, Karolinska University Hospital, Stockholm 171 76, Sweden.
authors.forename: Göran
authors.initials: G
authors.lastname: Carlsson
authors.affiliation: Karolinska Biomics Center, Department of Oncology and Pathology, Karolinska Institutet, Karolinska University Hospital, Stockholm 171 76, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Nordström
authors.affiliation: Karolinska Biomics Center, Department of Oncology and Pathology, Karolinska Institutet, Karolinska University Hospital, Stockholm 171 76, Sweden.
authors.forename: Janne
authors.initials: J
authors.lastname: Lehtiö
authors.affiliation: Department of Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, 751 85 Uppsala, Sweden.
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Fröjmark
authors.affiliation: Department of Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, 751 85 Uppsala, Sweden.
authors.forename: Niklas
authors.initials: N
authors.lastname: Dahl
authors.affiliation: Division of Molecular Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm 171 76, Sweden. Electronic address: bengt.fadeel@ki.se.
authors.forename: Bengt
authors.initials: B
authors.lastname: Fadeel
mesh_terms: D000111:Acetylcysteine / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing; D000975:Antioxidants / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D000075747:Baculoviral IAP Repeat-Containing 3 Protein; D020169:Caspases / Q000378:metabolism*; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic; D003374:Coumarins / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D005456:Fluorescent Dyes / Q000378:metabolism; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000378:metabolism; D019169:Jurkat Cells; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*; D044767:Ubiquitin-Protein Ligases; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000975:Antioxidants; D003374:Coumarins; D005456:Fluorescent Dyes; C104759:HAX1 protein, human; D051034:Inhibitor of Apoptosis Proteins; D016328:NF-kappa B; D009842:Oligopeptides; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D051636:X-Linked Inhibitor of Apoptosis Protein; C496866:XIAP protein, human; C102580:acetyl-aspartyl-glutamyl-valyl-aspartyl-amino-4-methylcoumarin; D014108:Tosylphenylalanyl Chloromethyl Ketone; C494833:BIRC2 protein, human; C494832:BIRC3 protein, human; D000075747:Baculoviral IAP Repeat-Containing 3 Protein; D044767:Ubiquitin-Protein Ligases; D011065:Poly(ADP-ribose) Polymerases; D020169:Caspases; D003545:Cysteine; D000111:Acetylcysteine
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