Different mode of cell death induced by calcium ionophore in human leukemia cell lines: possible role of constitutive endonuclease.
The mechanism of cell death induced by calcium ionophore, A23187, was investigated in six human leukemia cell lines. Following exposure to 1 microM A23187, the myelogenous cell lines (HL-60, U-937, KG-1) underwent apoptosis within 3 h as determined by their morphology and DNA fragmentation assay. In contrast, T-lymphoblastic leukemia cell lines (Molt-4, Molt-3, CEM) revealed necrotic cell death after 24 h of incubation. However, an initial rise of intracellular free calcium concentrations and growth inhibition after treatment with A23187 were similar in the two cell types. We further showed that an endonuclease capable of mediating internucleosomal DNA fragmentation was constitutively expressed in the cytosol but not in the nuclei of the myelogenous cell lines, although this endonuclease was not detected in either the nuclei or the cytosol of the T-lymphoblastic cell lines. The activation of the endonuclease in myelogenous cells is calcium-independent and has an optimal pH of 7.5-9. It is inhibited by 1 mM zinc ion or 300 microM aurintricarboxylic acid. We propose that this constitutive endonuclease may be related to the susceptibility of myelogenous leukemia cell lines to apoptotic cell death.
Matsubara
K
K
Department of Pediatrics, Faculty of Medicine, Kyoto University, Japan.
Kubota
M
M
Adachi
S
S
Kuwakado
K
K
Hirota
H
H
Wakazono
Y
Y
Akiyama
Y
Y
Mikawa
H
H
D017209:Apoptosis / Q000187:drug effects; D000001:Calcimycin / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects*; D004249:DNA Damage / Q000187:drug effects; D004720:Endonucleases / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D009336:Necrosis / Q000139:chemically induced; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000187:drug effects*
D000001:Calcimycin; D004720:Endonucleases; D002118:Calcium