The early phase of apoptosis in human neuroblastoma CHP100 cells is characterized by lipoxygenase-dependent ultraweak light emission.
Human neuroblastoma CHP100 cells were forced into apoptosis (programmed cell death, PCD) or necrosis by treatment with calcium chloride or sodium nitroprusside (a nitric oxide donor), respectively. Cellular luminescence, a marker of membrane lipid peroxidation, was increased by calcium but not by nitroprusside, and reached a maximum of 4-fold the control value 2 hours after treatment. The increase in luminescence was paralleled by increased 5-lipoxygenase (up to 250% of the control value) and decreased catalase (down to 50%) activity within the same time window. Consistently, incubation of CHP100 cells with inhibitors of 5-lipoxygenase (5,8,11,14-eicosatetraynoic acid and MK886) reduced light emission and PCD, whereas inhibition of catalase by 3-amino-1, 2,4-triazole enhanced both processes. Treatment of CHP100 cells with retinoic acid or cisplatin, unrelated PCD inducers reported to activate the lipoxygenase pathway, also gave enhanced light emission parallel to PCD increase. Altogether, these results suggest that cellular luminescence is an early marker of apoptotic, but not necrotic, program(s) involving generation of hydrogen peroxide and activation of 5-lipoxygenase.
Maccarrone
M
M
Department of Experimental Medicine and Biochemical Sciences, University of Rome Tor Vergata, Via di Tor Vergata 135, Rome, I-00133, Italy.
Salucci
M L
ML
Melino
G
G
Rosato
N
N
Finazzi-Agro
A
A
D015117:5,8,11,14-Eicosatetraynoic Acid / Q000494:pharmacology; D000640:Amitrole / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D002374:Catalase / Q000037:antagonists & inhibitors / Q000378:metabolism; D002945:Cisplatin / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D015289:Leukotrienes / Q000494:pharmacology; D015227:Lipid Peroxidation; D016859:Lipoxygenase Inhibitors; D008163:Luminescent Measurements; D009336:Necrosis; D009447:Neuroblastoma / Q000201:enzymology* / Q000473:pathology*; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D002722:Chlorobenzenes
D015117:5,8,11,14-Eicosatetraynoic Acid; D000640:Amitrole; D001094:Arachidonate 5-Lipoxygenase; D002374:Catalase; D002945:Cisplatin; D004791:Enzyme Inhibitors; D006861:Hydrogen Peroxide; D007211:Indoles; D015289:Leukotrienes; D016859:Lipoxygenase Inhibitors; D014212:Tretinoin; C060893:MK-886; D002722:Chlorobenzenes; C025348:arachidonic acid 5-hydroperoxide