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Nitric oxide-dependent apoptosis in ovarian carcinoma cell lines
J Rieder1, R Jahnke, M Schloesser
1Department of Anesthesia and Critical Care Medicine, Department of Obstetrics and Gynecology, University of Innsbruck, Anichstrasse 35, 6020, Innsbruck, Austria.
Objective:
In a recent study, we found different profiles of inducible nitric oxide synthase (iNOS) gene expression in the ovarian carcinoma cell lines OVCAR-3, HOC-7, and 2774 following stimulation by proinflammatory cytokines. The present study was performed to determine whether nitric oxide (NO) synthesis correlates with programmed cell death in these cells.
Methods:
NO-Dependent apoptosis was detected by DNA fragmentation analysis and fluorescence-activated cell sorter analysis.
Results:
NO formation in response to interferon gamma (IFN-gamma), interleukin-1beta (IL-1beta), and tumor necrosis factor alpha (TNF-alpha) was correlated with programmed cell death in the investigated cells. DNA fragmentation was most prominent in OVCAR-3 (34.17 +/- 1.81%), less pronounced in HOC-7 (12.86 +/- 0.45%), and undetectable in 2774 (4.54 +/- 0.40%) cells. The rate of apoptosis correlated with the amount of NO formation in cytokine-treated cells. Moreover, coincubation of OVCAR-3 and HOC-7 with the specific iNOS inhibitor aminoguanidine suppressed apoptosis induced by IFN-gamma, IL-1beta, and TNF-alpha.
Conclusion:
Our data indicate that in OVCAR-3 and HOC-7 cells, NO synthesis induced by IFN-gamma, IL-1beta, and TNF-alpha is correlated with the degree of apoptotic cell death. In clinical situations, this might in part explain the benefit of cytokine application in ovarian carcinoma patients (e.g., documented for IFN-gamma).
Insights
Nitric oxide (NO) synthesis correlates with programmed cell death in ovarian carcinoma cells. Cytokine-induced NO production in OVCAR-3 and HOC-7 cells promoted apoptosis, suggesting a role for NO in cancer therapy.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- Ovarian carcinoma cell lines (OVCAR-3, HOC-7, 2774) exhibit distinct inducible nitric oxide synthase (iNOS) gene expression profiles.
- Proinflammatory cytokines like interferon gamma (IFN-gamma), interleukin-1beta (IL-1beta), and tumor necrosis factor alpha (TNF-alpha) modulate iNOS expression.
Purpose of the Study:
- To investigate the correlation between nitric oxide (NO) synthesis and programmed cell death (apoptosis) in ovarian carcinoma cell lines.
- To determine the role of cytokine-induced NO in mediating apoptosis in these cells.
Main Methods:
- Apoptosis was assessed using DNA fragmentation analysis.
- Fluorescence-activated cell sorter (FACS) analysis was employed to quantify NO-dependent apoptosis.
- The effect of the iNOS inhibitor aminoguanidine on apoptosis was evaluated.
Main Results:
- NO formation in response to IFN-gamma, IL-1beta, and TNF-alpha correlated with programmed cell death.
- OVCAR-3 cells showed the highest DNA fragmentation (34.17%), followed by HOC-7 (12.86%), with minimal fragmentation in 2774 cells (4.54%).
- Aminoguanidine treatment suppressed cytokine-induced apoptosis in OVCAR-3 and HOC-7 cells.
Conclusions:
- NO synthesis induced by specific cytokines is linked to the extent of apoptotic cell death in OVCAR-3 and HOC-7 ovarian carcinoma cells.
- These findings suggest that NO plays a role in cytokine-mediated apoptosis in ovarian cancer.
- The observed correlation may partially explain the therapeutic benefits of cytokine application in ovarian carcinoma patients.