Pyrrolidinedithiocarbamate induces apoptosis in cerebellar granule cells: involvement of AP-1 and MAP kinases.
Pyrrolidinedithiocarbamate (PDTC) is a compound displaying antioxidant, pro-oxidant and metal chelator properties in different cell types. It has been described that PDTC may exert either anti-apoptotic or apoptotic activity. Moreover it is known that this agent regulates the activity of redox-sensitive transcription factors, such as AP-1 and NF-kappaB. Using cerebellar granule cells (CGCs), a well-described model of neuronal primary cultures, we investigated the effects of different concentrations of this compound on cell viability and the intracellular mechanisms involved. PDTC used at concentrations, as low as 1 microM, exerts cytotoxic effects on CGC through the activation of the apoptotic machinery with a maximal efficacy for concentration of 10 microM. The PDTC-dependent apoptosis is correlated to a biphasic and long-lasting increase of AP-1 binding to the DNA, apparently without affecting the NF-kappaB whose activity was reduced only at much higher concentrations (100 microM). PDTC treatment enhanced ERK phosphorylation (maximal effect 1h) and p38 phosphorylation (maximal effect 7h) that was accompanied by an increase of both mRNA and protein of c-Jun. In conclusion the results presented show that PDTC exerts apoptotic effects on CGC, that are correlated to the activation of stress-pathways, involving mainly AP-1 and MAPKs.
Porcile
Carola
C
Section of Pharmacology and Neuroscience, National Institute for Cancer Research (IST), CBA, Largo Rosanna Benzi 10, c/o Advanced Biotechnology Center (CBA), 16132, Genova, Italy.
Stanzione
Sabrina
S
Piccioli
Patrizia
P
Bajetto
Adriana
A
Barbero
Simone
S
Bisaglia
Michela
M
Bonavia
Rudy
R
Florio
Tullio
T
Schettini
Gennaro
G
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D006690:Bisbenzimidazole; D015153:Blotting, Western; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects*; D003594:Cytoplasmic Granules / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D024202:Electrophoretic Mobility Shift Assay; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D020928:Mitogen-Activated Protein Kinases / Q000187:drug effects* / Q000235:genetics; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000737:chemistry; D011759:Pyrrolidines / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013778:Tetrazolium Salts; D013844:Thiazoles; D013859:Thiocarbamates / Q000494:pharmacology*; D018808:Transcription Factor AP-1 / Q000187:drug effects*; D048051:p38 Mitogen-Activated Protein Kinases
D005456:Fluorescent Dyes; D016328:NF-kappa B; D009419:Nerve Tissue Proteins; D011759:Pyrrolidines; D013778:Tetrazolium Salts; D013844:Thiazoles; D013859:Thiocarbamates; D018808:Transcription Factor AP-1; C020972:pyrrolidine dithiocarbamic acid; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C022616:thiazolyl blue; D006690:Bisbenzimidazole