Zinc induces expression of the BH3-only protein PUMA through p53 and ERK pathways in SH-SY5Y neuroblastoma cells.
Accumulating evidence suggests that zinc (Zn2+) contributes to neuronal death in pathologic states such as ischemia. p53-upregulated modulator of apoptosis (PUMA), which is a BH3-only protein, is known to promote apoptosis through a tumor suppressor p53-dependent and -independent mechanism. In this study, we examined the effect of Zn2+ on the induction of the PUMA gene in human neuroblastoma SH-SY5Y cells. The expression of PUMA was induced by Zn2+ in a dose- and time-dependent manner. A reporter assay revealed that Zn2+ activated the PUMA promoter. In addition, the mutation of the p53 binding site in the PUMA promoter region reduced promoter activation by Zn2+. These findings suggest that p53 participates in Zn2+-induced PUMA expression. Furthermore, we also demonstrated here that Zn2+ stimulates the phosphorylation of ERK and that the MEK-ERK pathway inhibitor, U0126, suppressed Zn2+-induced PUMA expression. Taken together, these results indicate that Zn2+ regulates the induction of PUMA through p53 and ERK pathways.
Hara
Hirokazu
H
Laboratory of Clinical Pharmaceutics, Gifu Pharmaceutical University, 5-6-1 Mitahora-Higashi, Gifu 502-8585, Japan. harah@gifu-pu.ac.jp
Kamiya
Tetsuro
T
Adachi
Tetsuo
T
D051017:Apoptosis Regulatory Proteins / Q000502:physiology*; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000235:genetics*; D045744:Cell Line, Tumor; D004273:DNA, Neoplasm / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D016158:Genes, p53 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D009447:Neuroblastoma / Q000235:genetics*; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology; D015032:Zinc / Q000037:antagonists & inhibitors / Q000494:pharmacology*
D051017:Apoptosis Regulatory Proteins; C438378:BBC3 protein, human; D004273:DNA, Neoplasm; D004791:Enzyme Inhibitors; D011518:Proto-Oncogene Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D015032:Zinc