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Targeting hepatoma using nitric oxide donor strategies
Raúl González1, Gustavo Ferrín, Patricia Aguilar-Melero
1Department of Biochemistry and Molecular Biology, University of Córdoba, Córdoba, Spain.
Aims:
The study evaluated the role of increased intracellular nitric oxide (NO) concentration using NO donors or stably NO synthase-3 (NOS-3) overexpression during CD95-dependent cell death in hepatoma cells. The expression of cell death receptors and caspase activation, RhoA kinase activity, NOS-3 expression/activity, oxidative/nitrosative stress, and p53 expression were analyzed. The antitumoral activity of NO was also evaluated in the subcutaneous implantation of NOS-3-overexpressing hepatoma cells, as well NO donor injection into wild-type hepatoma-derived tumors implanted in xenograft mouse models.
Results:
NO donor increased CD95 expression and activation of caspase-8 and 3 in HepG2, Huh7, and Hep3B cells. NOS-3 overexpression increased oxidative/nitrosative stress, p53 and CD95 expression, cellular Fas-associated death domain (FADD)-like IL-1beta converting enzyme (FLICE) inhibitory protein long (cFLIP(L)) and its short isoform (cFLIP(S)) shift, and cell death in HepG2 (4TO-NOS) cells. The inhibition of RhoA kinase and p53 knockdown using RNA interference reduced cell death in 4TO-NOS cells. The supplementation with hydrogen peroxide (H(2)O(2)) increased NOS-3 activity and cell death in 4TO-NOS cells. NOS-3 overexpression or NO donor injection into hepatoma-derived tumors reduced the size and increased p53 and cell death receptor expression in nude mice.
Innovation And Conclusions:
The increase of intracellular NO concentration promoted oxidative and nitrosative stress, Rho kinase activity, p53 and CD95 expression, and cell death in cultured hepatoma cells. NOS-3-overexpressed HepG2 cells or intratumoral NO donor administration reduced tumor cell growth and increased the expression of p53 and cell death receptors in tumors developed in a xenograft mouse model.
Insights
Increasing nitric oxide (NO) in hepatoma cells enhances CD95-dependent cell death and reduces tumor growth. This study highlights NO
Area of Science:
- Hepatoma cell death mechanisms
- Nitric oxide signaling in cancer
- Tumor biology and therapy
Background:
- Hepatoma cells are a significant cause of cancer mortality.
- Nitric oxide (NO) plays a complex role in cell survival and death.
- Understanding NO's role in CD95-dependent apoptosis is crucial for cancer therapy.
Purpose of the Study:
- To investigate the effect of increased intracellular nitric oxide (NO) on CD95-dependent cell death in hepatoma cells.
- To analyze the impact of NO on cell death receptors, caspase activation, RhoA kinase activity, and p53 expression.
- To evaluate the antitumoral activity of NO in vivo using xenograft mouse models.
Main Methods:
- NO donors and NOS-3 overexpression were used to increase intracellular NO concentration.
- Analysis of cell death receptors, caspase activation, RhoA kinase activity, NOS-3 expression/activity, oxidative/nitrosative stress, and p53 expression.
- Xenograft mouse models with hepatoma cells (NOS-3 overexpressing or wild-type) were used to assess antitumoral effects.
Main Results:
- NO donors and NOS-3 overexpression increased CD95 expression and caspase activation in hepatoma cell lines.
- NOS-3 overexpression led to increased oxidative/nitrosative stress, p53 expression, and cell death.
- In vivo studies showed reduced tumor size and increased p53 and cell death receptor expression with NOS-3 overexpression or NO donor injection.
Conclusions:
- Increased intracellular NO concentration promotes oxidative/nitrosative stress, Rho kinase activity, p53 and CD95 expression, and cell death in hepatoma cells.
- NO-mediated mechanisms, including p53 and CD95 pathways, contribute to antitumoral effects.
- Intratumoral NO administration or NOS-3 overexpression demonstrates therapeutic potential against hepatoma in vivo.
