title: Adenosine- and 2-chloro-adenosine-induced cytopathic effects on myoblastic cells and myotubes: involvement of different intracellular mechanisms.
abstract: We recently suggested that, in muscular dystrophies, the excessive accumulation of adenosine as a result of an altered purine metabolism may contribute to progressive functional deterioration and muscle cell death. To verify this hypothesis, we have taken advantage of C2C12 myoblastic cells, which can be differentiated in vitro into multinucleated cells (myotubes). Exposure of both proliferating myoblasts and differentiated myotubes to adenosine or its metabolically-stable analog, 2-chloro-adenosine, resulted in apoptotic cell death and myotube disruption. Cytotoxicity by either nucleoside did not depend upon extracellular adenosine receptors, but, at least in part, by entry into cells via the membrane nitro-benzyl-thio-inosine-sensitive transporter. The adenosine kinase inhibitor, 5-iodotubercidin, prevented 2-chloro-adenosine-induced (but not adenosine-induced) effects, suggesting that an intracellular phosphorylation/activation reaction plays a key role in 2-chloro-adenosine-mediated cytotoxicity. Conversely, adenosine cytotoxicity was aggravated by the addition of homocysteine, suggesting that adenosine effects may be due to the accumulation of S-adenosyl-homocysteine, which blocks intracellular methylation-dependent reactions. Both nucleosides markedly disrupted the myotube structure via an effect on the actin cytoskeleton; however, also for myotubes, there were marked differences in the morphological alterations induced by these two nucleosides. These results show that adenosine and 2-chloro-adenosine induce apoptosis of myogenic cells via completely different metabolic pathways, and are consistent with the hypothesis that adenosine accumulation in dystrophic muscles may represent a novel pathogenetic pathway in muscle diseases.
authors.affiliation: Institute of Pharmacological Sciences, University of Milan, Via Balzaretti 9, 20133, Milan, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Ceruti
authors.forename: A M
authors.initials: AM
authors.lastname: Giammarioli
authors.forename: A
authors.initials: A
authors.lastname: Camurri
authors.forename: L
authors.initials: L
authors.lastname: Falzano
authors.forename: S
authors.initials: S
authors.lastname: Rufini
authors.forename: C
authors.initials: C
authors.lastname: Frank
authors.forename: C
authors.initials: C
authors.lastname: Fiorentini
authors.forename: W
authors.initials: W
authors.lastname: Malorni
authors.forename: M P
authors.initials: MP
authors.lastname: Abbracchio
mesh_terms: D015762:2-Chloroadenosine / Q000494:pharmacology*; D000111:Acetylcysteine / Q000494:pharmacology; D008841:Actin Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure; D000241:Adenosine / Q000378:metabolism* / Q000494:pharmacology; D000248:Adenosine Kinase / Q000037:antagonists & inhibitors; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016166:Free Radical Scavengers / Q000494:pharmacology; D006710:Homocysteine / Q000378:metabolism / Q000494:pharmacology; D007424:Intracellular Fluid / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D058915:Purinergic P1 Receptor Antagonists; D017382:Reactive Oxygen Species / Q000378:metabolism; D013868:Thioinosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014372:Tubercidin / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D058915:Purinergic P1 Receptor Antagonists; D017382:Reactive Oxygen Species; D006710:Homocysteine; D015762:2-Chloroadenosine; C034214:5-iodotubercidin; D013868:Thioinosine; D000248:Adenosine Kinase; C001789:4-nitrobenzylthioinosine; D000241:Adenosine; D014372:Tubercidin; D000111:Acetylcysteine