Characterization of catechol-thioether-induced apoptosis in human SH-SY5Y neuroblastoma cells.
Recent work has highlighted the involvement of a dopamine derivative, 5-S-cysteinyl-dopamine (CysDA), in neurodegeneration and apoptotic cell death. In this paper we study in further detail the apoptotic process activated by this catechol-thioether derivative of dopamine in SH-SY5Y neuroblastoma cells. CysDA activates a cascade of events by an initial perturbation of Calcium homeostasis in the cell. Cell treatment with the catechol-thioether induces an immediate rise in intracellular Ca(2+) concentration, as demonstrated by a shift in the indo-1 dye emission spectrum, and a sustained high calcium concentration at long times of incubation. Fluorescence microscopy data show that the treatment of cells induces mitochondrial transmembrane potential depolarization, a clear evidence of the onset of apoptotic process. Programmed cell death activation is also demonstrated by cytochrome c release from the mitochondria, by an increased activity of both caspase-8 and -9 and by the poly(ADP-ribose)polymerase (PARP-1) cleavage, yielding the typical 86 kDa fragment due to caspase-3 activity. Overall, our data support the hypothesis that CysDA may induce apoptotic death in neuronal cells, via an initial perturbation of calcium homeostasis in the cytosol.
Mosca
Luciana
L
Department of Biochemical Sciences, University Sapienza, Roma, Italy. luciana.mosca@uniroma1.it
Tempera
Italo
I
Lendaro
Eugenio
E
Di Francesco
Laura
L
d'Erme
Maria
M
D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002396:Catechols; D045744:Cell Line, Tumor; D003548:Cysteinyldopa / Q000494:pharmacology*; D045304:Cytochromes c / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000187:drug effects; D009447:Neuroblastoma; D009474:Neurons / Q000187:drug effects*; D011065:Poly(ADP-ribose) Polymerases / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology
D002396:Catechols; D003548:Cysteinyldopa; D045304:Cytochromes c; D011065:Poly(ADP-ribose) Polymerases; C034221:catechol; D002118:Calcium