title: Artesunate induces ROS- and p38 MAPK-mediated apoptosis and counteracts tumor growth in vivo in embryonal rhabdomyosarcoma cells.
abstract: Rhabdomyosarcoma represents about 50% of soft-tissue sarcomas and 10% of malignant solid tumors in childhood. Embryonal rhabdomyosarcoma (ERMS) is the most frequent subtype, suggested to have an origin in muscle precursor cells that fail to exit the cell cycle and terminally differentiate mainly because of overexpression of the transcription factor, PAX7, which sustains proliferation, migration and invasiveness in ERMS cells. Artesunate (ARS) is a semi-synthetic derivative of artemisinin (ART), a natural compound well known as an antimalarial drug. However, ART and its derivatives have been found efficacious even as anticancer drugs that induce cell cycle arrest and/or apoptosis in several kinds of cancer. Here, we show that ARS dose-dependently induces DNA damage and apoptosis in ERMS cell lines. Production of reactive oxygen species (ROS) and activation of p38 MAPK have a central role in triggering ARS-mediated apoptosis in ERMS cells; indeed either the antioxidant, N-acetylcysteine or the p38 MAPK inhibitor, SB203580, protects ERMS cells from ARS-induced apoptosis. Moreover, ARS treatment in ERMS cells ROS-dependently induces the expression of the myo-miRs, miR-133a and miR-206, which are down-regulated in RMS, and reduces PAX7 protein levels. Finally, ARS upregulates the expression of the adhesion molecules, NCAM and integrin β1, and reduces migration and invasiveness of ERMS cells in vitro, and ARS treatment reduces of about 50% the growth of ERMS xenografts in vivo. Our results are the first evidence of efficacy of ART derivatives in restraining ERMS growth in vivo, and suggest ARS as a potential candidate for therapeutic treatment of ERMS.
authors.affiliation: Department of Experimental Medicine, University of Perugia 06132 Perugia, Italy, Interuniversity Institute of Myology (IIM), Padova, Italy and.
authors.forename: Sara
authors.initials: S
authors.lastname: Beccafico
authors.affiliation: Department of Experimental Medicine, University of Perugia 06132 Perugia, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Morozzi
authors.affiliation: Department of Medicine, University of Perugia 06132 Perugia, Italy.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Marchetti
authors.affiliation: Department of Medicine, University of Perugia 06132 Perugia, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Riccardi
authors.affiliation: Department of Experimental Medicine, University of Perugia 06132 Perugia, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Sidoni
authors.affiliation: Department of Experimental Medicine, University of Perugia 06132 Perugia, Italy, Interuniversity Institute of Myology (IIM), Padova, Italy and guglielmo.sorci@unipg.it rosario.donato@unipg.it.
authors.forename: Rosario
authors.initials: R
authors.lastname: Donato
authors.affiliation: Department of Experimental Medicine, University of Perugia 06132 Perugia, Italy, Interuniversity Institute of Myology (IIM), Padova, Italy and guglielmo.sorci@unipg.it.
authors.forename: Guglielmo
authors.initials: G
authors.lastname: Sorci
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D037621:Artemisinins / Q000494:pharmacology* / Q000627:therapeutic use; D000077332:Artesunate; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D004249:DNA Damage / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D019012:Integrin beta1 / Q000096:biosynthesis; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D019006:Neural Cell Adhesion Molecules / Q000096:biosynthesis; D051763:PAX7 Transcription Factor / Q000096:biosynthesis; D011725:Pyridines / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D018233:Rhabdomyosarcoma, Embryonal / Q000188:drug therapy* / Q000473:pathology*; D014183:Transplantation, Heterologous; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D037621:Artemisinins; D007093:Imidazoles; D019012:Integrin beta1; C548234:MIRN133 microRNA, human; C546087:MIRN206 microRNA, human; D035683:MicroRNAs; D019006:Neural Cell Adhesion Molecules; D051763:PAX7 Transcription Factor; C497321:PAX7 protein, human; D011725:Pyridines; D017382:Reactive Oxygen Species; D000077332:Artesunate; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580; D000111:Acetylcysteine