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Upregulation and activation of caspase-3 or caspase-8 and elevation of intracellular free calcium mediated apoptosis of indomethacin-induced K562 cells / 中华医学杂志(英文版)
Chinese Medical Journal ; (24): 978-984, 2004.
Article in English | WPRIM | ID: wpr-284865
ABSTRACT
<p><b>BACKGROUND</b>A nonsteroidal anti-inflammatory drug, indomethacin, has been shown to have anti-leukemic activity and induce leukemic cell apoptosis. This study was to elucidate the mechanism of indomethacin-induced K562 cell apoptosis.</p><p><b>METHODS</b>K562 cells were grown in RPMI 1640 medium and treated with different doses of indomethacin (0 micromol/L, 100 micromol/L, 200 micromol/L, 400 micromol/L, 800 micromol/L) for 72 hours. The cells were harvested, and cell viability or apoptosis was analyzed using MTT assay and AO/EB stain, combining laser scanning confocal microscopy (LSCM) technique separately. For the localization and distribution of intracellular caspase-3 or caspase-8 protein, immunofluorescence assay was carried out. To reveal the activation of caspase-3 or caspase-8 in indomethacin-treated cells, Western blot detection was used. The change in intracellular free calcium was determined by Fluo-3/Am probe labeling combined with LSCM.</p><p><b>RESULTS</b>Indomethacin could lead to K562 cell apoptosis and inhibit cell viability in a concentration-dependent manner. An increased expression of intracellular caspase-3 or caspase-8 was observed at higher doses of indomethacin (400 - 800 micromol/L). Western blot results showed upregulation and activation in both caspase-3 and caspase-8 protein. Under indomethacin intervention, the levels of intracellular free calcium showed a significant increase. Blocking the activity of cyclooxygenase did not abolish the effects of indomethacin on K562 cell apoptosis.</p><p><b>CONCLUSIONS</b>Activation and upregulation of caspase-3 or caspase-8 protein were responsible for Indomethacin-induced K562 cell apoptosis. Variation of intracellular free calcium might switch on the apoptotic pathway and the proapoptotic effect of indomethacin might be cyclooxygenase-independent.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Pharmacology / Gene Expression Regulation, Enzymologic / Indomethacin / Calcium / Cyclooxygenase Inhibitors / Apoptosis / K562 Cells / Caspases / Enzyme Activation / Caspase 3 Limits: Humans Language: English Journal: Chinese Medical Journal Year: 2004 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Pharmacology / Gene Expression Regulation, Enzymologic / Indomethacin / Calcium / Cyclooxygenase Inhibitors / Apoptosis / K562 Cells / Caspases / Enzyme Activation / Caspase 3 Limits: Humans Language: English Journal: Chinese Medical Journal Year: 2004 Type: Article