Thiamine deficiency caused by thiamine antagonists triggers upregulation of apoptosis inducing factor gene expression and leads to caspase 3-mediated apoptosis in neuronally differentiated rat PC-12 cells.
Recent evidence suggests that alterations in oxidative metabolism induced by thiamine deficiency lead to neuronal cell death. However, the molecular mechanisms underlying this process are still under extensive investigation. Here, we report that rat pheochromocytoma PC-12 cells differentiated in the presence of NGF into neurons undergo apoptosis due to thiamine deficiency caused by antagonists of thiamine - amprolium, pyrithiamine and oxythiamine. Confocal laser scanning fluorescence microscopy revealed that annexin V binds to PC-12 cells in presence of thiamine antagonists after 72 h incubation. Results also show that thiamine antagonists trigger upregulation of gene expression of mitochondrial-derived apoptosis inducing factor, DNA fragmentation, cleavage of caspase 3 and translocation of active product to the nucleus. We therefore propose that apoptosis induced by amprolium, pyrithiamine or oxythiamine occurs via the mitochondria-dependent caspase 3-mediated signaling pathway. In addition, our data indicate that pyrithiamine and oxythiamine are more potent inducers of apoptosis than amprolium.
Chornyy
Sergiy
S
Department of Coenzyme Biochemistry, Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv, Ukraine. sch_1982@yahoo.com
Parkhomenko
Julia
J
Chorna
Nataliya
N
D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000670:Amprolium / Q000494:pharmacology; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D051033:Apoptosis Inducing Factor / Q000235:genetics*; D053148:Caspase 3 / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D020932:Nerve Growth Factor / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010119:Oxythiamine / Q000494:pharmacology; D016716:PC12 Cells; D011485:Protein Binding / Q000187:drug effects; D011745:Pyrithiamine / Q000494:pharmacology; D051381:Rats; D013831:Thiamine / Q000037:antagonists & inhibitors*; D013832:Thiamine Deficiency / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D015854:Up-Regulation
D017304:Annexin A5; D051033:Apoptosis Inducing Factor; D010119:Oxythiamine; D011745:Pyrithiamine; D020932:Nerve Growth Factor; C505410:Casp3 protein, rat; D053148:Caspase 3; D000670:Amprolium; D013831:Thiamine