title: Lobatin B inhibits NPM/ALK and NF-κB attenuating anaplastic-large-cell-lymphomagenesis and lymphendothelial tumour intravasation.
abstract: An apolar extract of the traditional medicinal plant Neurolaena lobata inhibited the expression of the NPM/ALK chimera, which is causal for the majority of anaplastic large cell lymphomas (ALCLs). Therefore, an active principle of the extract, the furanoheliangolide sesquiterpene lactone lobatin B, was isolated and tested regarding the inhibition of ALCL expansion and tumour cell intravasation through the lymphendothelium. ALCL cell lines, HL-60 cells and PBMCs were treated with plant compounds and the ALK inhibitor TAE-684 to measure mitochondrial activity, proliferation and cell cycle progression and to correlate the results with protein- and mRNA-expression of selected gene products. Several endpoints indicative for cell death were analysed after lobatin B treatment. Tumour cell intravasation through lymphendothelial monolayers was measured and potential causal mechanisms were investigated analysing NF-κB- and cytochrome P450 activity, and 12(S)-HETE production. Lobatin B inhibited the expression of NPM/ALK, JunB and PDGF-Rβ, and attenuated proliferation of ALCL cells by arresting them in late M phase. Mitochondrial activity remained largely unaffected upon lobatin B treatment. Nevertheless, caspase 3 became activated in ALCL cells. Also HL-60 cell proliferation was attenuated whereas PBMCs of healthy donors were not affected by lobatin B. Additionally, tumour cell intravasation, which partly depends on NF-κB, was significantly suppressed by lobatin B most likely due to its NF-κB-inhibitory property. Lobatin B, which was isolated from a plant used in ethnomedicine, targets malignant cells by at least two properties: I) inhibition of NPM/ALK, thereby providing high specificity in combating this most prevalent fusion protein occurring in ALCL; II) inhibition of NF-κB, thereby not affecting normal cells with low constitutive NF-κB activity. This property also inhibits tumour cell intravasation into the lymphatic system and may provide an option to manage this early step of metastatic progression.
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, A-1090 Vienna, Austria; Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
authors.forename: Izabella
authors.initials: I
authors.lastname: Kiss
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, A-1090 Vienna, Austria.
authors.forename: Christine
authors.initials: C
authors.lastname: Unger
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
authors.forename: Chi Nguyen
authors.initials: CN
authors.lastname: Huu
authors.affiliation: Department of Pharmacognosy, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
authors.forename: Atanas Georgiev
authors.initials: AG
authors.lastname: Atanasov
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, A-1090 Vienna, Austria.
authors.forename: Nina
authors.initials: N
authors.lastname: Kramer
authors.affiliation: Department of Clinical Pharmacy and Diagnostics, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria; Department of Preclinic, Faculty of Medicine, Mahasarakham University, Mahasarakham 44000, Thailand.
authors.forename: Waranya
authors.initials: W
authors.lastname: Chatruphonprasert
authors.affiliation: Department of Clinical Pharmacy and Diagnostics, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
authors.forename: Stefan
authors.initials: S
authors.lastname: Brenner
authors.affiliation: Department of Pharmacognosy, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
authors.forename: Ruxandra
authors.initials: R
authors.lastname: McKinnon
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
authors.forename: Andrea
authors.initials: A
authors.lastname: Peschel
authors.affiliation: Department of Pharmacognosy, University of Szeged, Eotvos Str. 6, H-6720 Szeged, Hungary.
authors.forename: Andrea
authors.initials: A
authors.lastname: Vasas
authors.affiliation: Department of Pharmacognosy, University of Szeged, Eotvos Str. 6, H-6720 Szeged, Hungary.
authors.forename: Ildiko
authors.initials: I
authors.lastname: Lajter
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
authors.forename: Renate
authors.initials: R
authors.lastname: Kain
authors.affiliation: Department of Medical and Chemical Laboratory Diagnostics, Medical University of Vienna, Waehringer Guertel 18-20, Austria.
authors.forename: Philipp
authors.initials: P
authors.lastname: Saiko
authors.affiliation: Department of Medical and Chemical Laboratory Diagnostics, Medical University of Vienna, Waehringer Guertel 18-20, Austria.
authors.forename: Thomas
authors.initials: T
authors.lastname: Szekeres
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria; Ludwig Boltzmann Institute for Cancer Research, LBI-CR, Waehringerstrasse 13a, 1090 Vienna, Austria; Unit of Pathology of Laboratory Animals, University of Veterinary Medicine Vienna, 1210 Vienna, Austria.
authors.forename: Lukas
authors.initials: L
authors.lastname: Kenner
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
authors.forename: Melanie R
authors.initials: MR
authors.lastname: Hassler
authors.affiliation: Institute for Ethnobiology, Playa Diana, San José, Petén, Guatemala.
authors.forename: Rene
authors.initials: R
authors.lastname: Diaz
authors.affiliation: Institute for Ethnobiology, Playa Diana, San José, Petén, Guatemala.
authors.forename: Richard
authors.initials: R
authors.lastname: Frisch
authors.affiliation: Department of Pharmacognosy, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
authors.forename: Verena M
authors.initials: VM
authors.lastname: Dirsch
authors.affiliation: Department of Clinical Pharmacy and Diagnostics, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
authors.forename: Walter
authors.initials: W
authors.lastname: Jäger
authors.affiliation: Department of Vascular Biology and Thrombosis Research, Center of Biomolecular Medicine and Pharmacology, Medical University of Vienna, Schwarzspanierstraße 17, A-1090 Vienna, Austria.
authors.forename: Rainer
authors.initials: R
authors.lastname: de Martin
authors.affiliation: Institute of Pharmaceutical Sciences, University of Graz, Schubertstraße 1, A-8010 Graz, Austria.
authors.forename: Valery N
authors.initials: VN
authors.lastname: Bochkov
authors.affiliation: Institute of Pharmaceutical Biology and Biotechnology, Heinrich-Heine-University Düsseldorf, Universitätsstrasse 1, D-40225 Düsseldorf, Germany.
authors.forename: Claus M
authors.initials: CM
authors.lastname: Passreiter
authors.affiliation: Department of Medicine I, Division: Institute of Cancer Research, Comprehensive Cancer Center, Medical University Vienna, Borschkegasse 8a, A-1090 Vienna, Austria.
authors.forename: Barbara
authors.initials: B
authors.lastname: Peter-Vörösmarty
authors.affiliation: Department of Medicine I, Comprehensive Cancer Center, Medical University Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Mader
authors.affiliation: Department of Medicine I, Division: Institute of Cancer Research, Comprehensive Cancer Center, Medical University Vienna, Borschkegasse 8a, A-1090 Vienna, Austria.
authors.forename: Michael
authors.initials: M
authors.lastname: Grusch
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, A-1090 Vienna, Austria.
authors.forename: Helmut
authors.initials: H
authors.lastname: Dolznig
authors.affiliation: Department of Pharmacognosy, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Kopp
authors.affiliation: Department of Pharmacodynamics and Biopharmacy, University of Szeged, H-6720 Szeged, Hungary.
authors.forename: Istvan
authors.initials: I
authors.lastname: Zupko
authors.affiliation: Department of Pharmacognosy, University of Szeged, Eotvos Str. 6, H-6720 Szeged, Hungary.
authors.forename: Judit
authors.initials: J
authors.lastname: Hohmann
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria. Electronic address: georg.krupitza@meduniwien.ac.at.
authors.forename: Georg
authors.initials: G
authors.lastname: Krupitza
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D019659:Asteraceae / Q000737:chemistry*; D015153:Blotting, Western; D020169:Caspases / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics / Q000378:metabolism; D004729:Endothelium, Lymphatic / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D017728:Lymphoma, Large-Cell, Anaplastic / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D009361:Neoplasm Invasiveness; D010936:Plant Extracts / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012717:Sesquiterpenes / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D000972:Antineoplastic Agents, Phytogenic; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016328:NF-kappa B; D010936:Plant Extracts; D016755:Proto-Oncogene Proteins c-jun; D012333:RNA, Messenger; D012717:Sesquiterpenes; C000595662:lobatin B; C107023:p80(NPM-ALK) protein; D011505:Protein-Tyrosine Kinases; D020169:Caspases
keywords: 3D-compound testing; ALCL; Lobatin; Lymphendothelial intravasation; NPM/ALK