Acetaminophen potentiates staurosporine-induced death in a human neuroblastoma cell line
I Posadas1, V Vellecco, P Santos
1Unidad Asociada Neurodeath, UCLM-CSIC, Universidad de Castilla-La Mancha, Avda. de Almansa, Albacete, Spain.
Background And Purpose:
Neuroblastoma is the most common solid tumour in infants characterized by a high resistance to apoptosis. Recently, the cyclo-oxygenase pathway has been considered a potential target in the treatment of different kinds of tumours. The aim of the present work was to investigate a possible relationship between cyclo-oxygenase pathway and stauroporine-induced apoptosis in the neuroblastoma cell line SH-SY5Y.
Experimental Approach:
Cellular viability was measured by release of LDH. DNA fragmentation was visualized by electrophoresis on agarose gel containing ethidium bromide. Cyclo-oxygenase activity was measured in microsomal fractions obtained from cells by quantification of its final product PGE2 by RIA. Caspase-3 activity was measured fluorimetrically and Western blot analysis was performed to assess cytochrome c expression.
Key Results:
We have found that staurosporine (500 nM) induced cellular death in a time-dependent manner in SH-SY5Y human neuroblastoma cells. Cyclo-oxygenase enzymatic activity was present in SH-SY5Y human neuroblastoma cells under basal conditions and pharmacological experiments using COX inhibitors indicate that cyclo-oxygenase-1 and cyclo-oxygenase-3 are the active isoforms in these cells. Co-incubation of SH-SY5Y cells with staurosporine (500 nM) and acetaminophen for 24 h potentiated staurosporine-mediated cellular death in a concentration-dependent manner. This process is mediated by an increase in cytochrome c release and caspase 3 activation and is prevented by N-acetylcysteine or the superoxide dismutase mimetic, MnTBAP.
Conclusions And Implications:
Acetaminophen potentiates staurosporine-mediated neuroblastoma cell death. The mechanism of action of acetaminophen seems to be related to production of reactive oxygen species and decreased intracellular glutathione levels.
Insights
Acetaminophen enhances staurosporine-induced apoptosis in neuroblastoma cells by increasing reactive oxygen species. This finding suggests a novel therapeutic approach for neuroblastoma, a common infant cancer.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Neuroblastoma, a common infant solid tumor, exhibits significant resistance to apoptosis.
- The cyclo-oxygenase (COX) pathway is increasingly recognized as a potential therapeutic target in various cancers.
- Understanding apoptosis resistance mechanisms is crucial for developing effective neuroblastoma treatments.
Purpose of the Study:
- To investigate the relationship between the cyclo-oxygenase pathway and staurosporine-induced apoptosis in the SH-SY5Y neuroblastoma cell line.
- To explore the potential role of acetaminophen in modulating apoptosis in neuroblastoma cells.
Main Methods:
- Cell viability assessed via lactate dehydrogenase (LDH) release.
- DNA fragmentation visualized using agarose gel electrophoresis.
- Cyclo-oxygenase activity measured by prostaglandin E2 (PGE2) quantification; caspase-3 activity and cytochrome c expression analyzed via fluorimetry and Western blot.
Main Results:
- Staurosporine induced time-dependent cell death in SH-SY5Y cells.
- Cyclo-oxygenase-1 and cyclo-oxygenase-3 were identified as the active isoforms in these cells.
- Acetaminophen potentiated staurosporine-mediated cell death, linked to increased cytochrome c release and caspase-3 activation, and was mitigated by N-acetylcysteine or MnTBAP.
Conclusions:
- Acetaminophen enhances staurosporine-induced apoptosis in neuroblastoma cells.
- The mechanism involves reactive oxygen species production and reduced intracellular glutathione.
- This suggests a potential therapeutic strategy involving acetaminophen for neuroblastoma treatment.
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